CN107405409A

CN107405409A - CNP prodrugs

Info

Publication number: CN107405409A
Application number: CN201680014717.9A
Authority: CN
Inventors: K·斯普罗格; U·赫泽尔; H·拉乌; T·韦格; F·佛汀各; F·克利曼; N·卡鲁扎; A·伯恩哈德; A·布巴; T·伍兹
Original assignee: Ascendis Pharma AS
Current assignee: Ascendis Pharma AS
Priority date: 2015-01-09
Filing date: 2016-01-08
Publication date: 2017-11-28
Anticipated expiration: 2036-01-08
Also published as: SG11201705521YA; US20190255183A1; CN107405409B; US20230116746A1; US20170368189A1; AU2021203717A1; JP2024016208A; MX2022005466A; RU2017128291A3; KR102599552B1; JP2018502868A; JP2021008515A; AU2016205968A1; IL278970B1; MX2017008915A; AU2023204227A1; SG10202011002WA; US20210069339A1; WO2016110577A1; IL278970A

Abstract

The present invention relates to the prodrug of C types natriuretic peptide (CNP), the pharmaceutical composition comprising the CNP prodrugs and their purposes.In one embodiment, the CNP prodrugs are that CNP peptides pass through reversible connector and the conjugate of PEG.

Description

CNP prodrugs

The present invention relates to CNP prodrugs, its pharmaceutically acceptable salt, comprising the CNP prodrugs or its is pharmaceutically acceptable Salt pharmaceutical composition and their purposes.

Gain-of-function mutation in FGFR3 causes achondroplasia (achondroplasia, ACH), cartilage development Bad (hypochondroplasia, HCH) and thanatophoric dysplasia (thanatophoric dysplasia, TD).Due to The increased signal transduction of fibroblast growth factor receptor3 (FGFR3), these illnesss are characterized in out-of-proportion limb root Type (rhizomelic) nanism and order of severity difference, its range from mild (HCH) arrive serious (ACH) and lethal (TD).FGFR3 is the key regulator of endochrondral bone growth, sends signal by several intracellular pathways, including signal turns Guide and those of transcription activator (STAT) and inhibition of mitogen-activated protein kinase (MAPK).FGFR3 constitutive activations damage The propagation and terminal differentiation of growth plate chondrocyte and the synthesis of extracellular matrix.FGFR3 is activated and STAT and MAPK paths Phosphorylation increase it is relevant.MAPK signal paths are adjusted by c-type natriuretic peptide (CNP).CNP and its acceptor natriuretic peptide receptor B (NPR- B combination) suppresses FGFR3 downstream signal transductions, so as to trigger endochondra1 growth and bone undue growth, is such as over-expressing What is observed in CNP mouse and the mankind.Excessive generations of the CNP in cartilage passes through intravenous (iv) and is transfused continuous conveying CNP makes the nanism normalization of the mouse of achondroplasia, shows CNP in the plan that the administration of physiological levels is treatment ACH Slightly.

However, it is contemplated that its half-life short (2 minutes after intravenous (iv) administration), CNP is as therapeutic agent in pediatric population In it is challenging because it needs continuous infusion.Further, since CNP is inactivated extensively in hypodermis, it is therefore desirable to quiet It is transfused in arteries and veins.

Potter (FEBS Journal 278 (2011) 1808-1817) describes CNP and occurred by two kinds of degradation pathways Remove：Receptor-mediated degraded and the degraded by extracellular protease.The work that CNP passes through neutral endopeptidase 24.11 (NEP) Removed with degraded, and by natriuretic peptide removing acceptor NPR-C systemic circulation, NPR-C is combined with CNP and is deposited into lyase In body, CNP is degraded in lysosome.

The ability for describing individual organ by extracting ratio and molecule being removed from circulation, extraction ratio is by from artery concentration In subtract venous concentration and calculate the value divided by the arterial blood concentration of molecule.This so-called A/V differences quantify organ How effectively remove or degrade molecule of interest.In the mankind, CNP A/V gradients be for kidney, liver and lung tissue it is negative, Degraded with the CNP occurred in these tissues consistent.

Reduce to degrade by one or two in these removing approach and will be helpful to the half-life period for extending CNP.

Because the size of its avtive spot chamber is limited, preferred substrates of the identification less than about 3kDa of NEP.US 8,377,884B2 CNP variant is described, it is optionally forever conjugated to increase the resistance to NEP cuttings with PEG polymer.However it has been found that to open country Raw type CNP addition even as low as 0.6kDa PEG reduce CNP activity, and are greater than about 2 or 3kDa to CNP or the addition of its variant PEG reduce CNP functional activity in a manner of size relies on.Therefore, the PEG for being more than 2 to 3kDa to be degraded for reducing NEP Along with the forfeiture of activity, this may reduce the treatment potentiality of these molecules for the connection of molecule.

In addition to negatively affecting the activity of peptide, PEG or another macromoleculars and the conjugated of CNP may also prevent effectively to divide Cloth is to growth plate.(the Anat Rec A Discov Mol Cell Evol Biol.2006January such as Farnum；288(1): 91-103) prove that distribution of the molecule from systemic blood vessels system to growth plate is size-dependent, and small molecule (highest It can 10kDa) be distributed to growth plate, and more than 40kDa molecular dimension entry deterrence growth plate.

International application WO 2009/156481 is related to BNP reversible PEG- conjugates, and the BNP terms are defined as including sharp sodium All members of peptide family.This application is concerned only with the Cardiovascular of this kind of peptide by natriuretic peptide receptor A (NPR-A) mediations. It is soft on the chondrosin long slab by activating natriuratic peptide receptor B (NPR-B) mediations that WO 2009/156481A1 fail open CNP The specific nature that growth, propagation and the differentiation of osteocyte are adjusted.

Establishment NEP is described in The American Journal of Human Genetics 91,1108-1114 to resist Property CNP molecules simultaneously can carry out the distinct methods of subcutaneous administration.BMN-111 is the recombined human c-type natriuretic peptide (CNP) through modification, 17 amino acid have wherein been added to form the CNP pharmacology analogs of 39 amino acid.BMN-111 simulates CNP in growth plate On pharmacological activity, and due to neutrality-endopeptidase (NEP) resistance and with extend half-life period, it is allowed to once a day Subcutaneously (SC) is applied.Because BMN-111 is non-naturally occurring peptide, compared with native peptides, inducing the risk of immune response increases, And as Martz " described in sFGFR for achondroplasia " (SciBx, Biocenture in October, 2013), The immune response to BMN-111 is observed in zooscopy, the antibody that the pharmacological activity for not influenceing medicine be present.However, BMN- 111 only have the half-life period of 20 minutes, related to the short duration exposed to effective levels of drugs when daily apply.

In order to increase to the exposure of Effective Dose Level, the dosage of the medicine with CNP activity can be increased.Due to sharp sodium Peptide is the hormone family that possible influence blood volume and blood pressure, and the increase of dosage may be relevant with cardiovascular adverse effects.BMN-111 Research in animals and humans shows, with the increase of dosage, arterial pressure declines and heart rate increase.Up to 15 μ g/kg's BMN-111 dosage is relevant with the slight low blood pressure of healthy volunteer.Therefore, the dosage with medicine active CNP is increased to increase Medicine exposure may be related to unacceptable cardiovascular side effects.

In a word, it is necessary to more convenient and/or effective CNP treatments.

It is therefore an object of the present invention to disadvantages mentioned above is overcome at least in part.

The purpose realizes that wherein prodrug has formula (Ia) or (Ib) by CNP prodrugs or its pharmaceutically acceptable salt Structure

Wherein

- D is CNP parts (moiety)；

-L¹- it is reversible prodrug connector part；

-L²- it is single chemical bond or interval body portion；

- Z is water-solubility carrier part；

X is to be selected from following integer：1st, 2,3,4,5,6,7,8,9,10,11,12,13,14,15 or 16；And

Y is to be selected from following integer：1st, 2,3,4 and 5.

On the other hand, the present invention relates to CNP prodrugs or its pharmaceutically acceptable salt, it includes conjugate D-L, wherein

- D is CNP parts；And

- L includes reversible prodrug connector part-L¹-；

Wherein-L¹- by-L²- Z ' substitutes and is optionally further substituted；Wherein

-L²- it is single chemical bond or interval body portion；And

- Z ' is water insoluble carrier part.

It should be understood that multiple-L²-L¹- D parts are connected with water insoluble carrier-Z '.

It has surprisingly been found that the CNP prodrugs and its pharmaceutically acceptable salt of the present invention provide CNP and prolonged in blood flow Long circulation time, it causes the friendly method of application of more convenient and patient, such as SC is noted once in a week or up to monthly Penetrate.Meanwhile unmodified CNP is discharged, it ensures activating agent effectively distribution to growth plate.Due to medical instrument before the CNP of the present invention There is low-residual active, i.e., combined with NPR-B, the risk of cardiovascular side effects such as low blood pressure significantly reduces.

In addition it has surprisingly been found that the compound acquisition of the present invention is more more stable than what is observed after daily bolus infusion Blood level, its closer to simulation endogenous CNP physiology exposure.These more stable blood levels are applied to various doses Amount scheme, such as applied suitable for daily；Per once two days, once every three days, per once four days, per once five days, every six days Once；Apply weekly；Double weeks apply and monthly applied.

In addition CNP sustained release, such as the controlled release durg delivery system ratio of the prodrug from the present invention are surprisingly found out that Inject once a day more effective.

In the present invention, using the term with following implication.

As used herein, term " CNP " refers to all CNP polypeptides, is preferred from mammalian species, more preferably comes from People and mammalian species, more preferably from people and mouse species, and it is their variant, analog, ortholog thing, same It is thing (homolog) and derivative and fragment, it is characterised in that the growth of regulation chondrosin long slab cartilage cell, breed and divide Change.Preferably, term " CNP " refers to SEQ ID NO:1 CNP polypeptides and its there is essentially identical bioactivity to adjust The growth of chondrosin long slab cartilage cell, variant, homologue and the derivative of propagation and differentiation.It is highly preferred that term " CNP " is Refer to SEQ ID NO:1 polypeptide.It is also preferred that term " CNP " refers to SEQ ID NO:24, i.e., it is made up of 38 amino acid CNP parts, and its show substantially the same bioactivity adjust chondrosin long slab cartilage cell growth, propagation and Variant, homologue and the derivative of differentiation.

SEQ ID NO:1 has following sequence：

GLSKGCFGLKLDRIGSMSGLGC,

Wherein 6 are connected with the cysteine of 22 by disulphide bridges, as shown in Figure 1.

SEQ ID NO:24 have following sequence：

LQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC,

Wherein 22 are connected with the cysteine of 38 by disulphide bridges.

All CNP that term " CNP " is additionally included in disclosed in WO 2009/067639A2 and WO 2010/135541A2 become Body, analog, ortholog thing, homologue and its derivative and fragment, are incorporated herein by reference.

Therefore, term " CNP " also preferably refers to following peptide sequence：

SEQ ID NO：2(CNP-53)：

DLRVDTKSRAAWARLLQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC；

SEQ ID NO：3(G-CNP-53)：

GDLRVDTKSRAAWARLLQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC；

SEQ ID NO：4(M-CNP-53)：

MDLRVDTKSRAAWARLLQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC；

SEQ ID NO：5(P-CNP-53)：

PDLRVDTKSRAAWARLLQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC；

SEQ ID NO.6(CNP-53M48N)：

DLRVDTKSRAAWARLLQEHPNARKYKGANKKGLSKGCFGLKLDRIGSNSGLGC；

SEQ ID NO：7(CNP-53Δ15-31)：

DLRVDTKSRAAWARGLSKGCFGLKLDRIGSMSGLGC；

SEQ ID NO：8(CNP-52)：

LRVDTKSRAAWARLLQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC；

SEQ ID NO：9(CNP-51)：

RVDTKSRAAWARLLQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC；

SEQ ID NO：10(CNP-50)：

VDTKSRAAWARLLQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC；

SEQ ID NO：11(CNP-49)：

DTKSRAAWARLLQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC；

SEQ ID NO：12(CNP-48)：

TKSRAAWARLLQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC；

SEQ ID NO：13(CNP-47)：

KSRAAWARLLQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC；

SEQ ID NO：14(CNP-46)：

SRAAWARLLQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC；

SEQ ID NO：15(CNP-45)：

RAAWARLLQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC；

SEQ ID NO：16(CNP-44)：

AAWARLLQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC；

SEQ ID NO：17(CNP-44Δ14-22)：

AAWARLLQEHPNAGLSKGCFGLKLDRIGSMSGLGC；

SEQ ID NO：18(CNP-44Δ15-22)：

AAWARLLQEHPNARGLSKGCFGLKLDRIGSMSGLGC；

SEQ ID NO：19(CNP-43)：

AWARLLQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC；

SEQ ID NO：20(CNP-42)：

WARLLQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC；

SEQ ID NO：21(CNP-41)：

ARLLQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC；

SEQ ID NO：22(CNP-40)：

RLLQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC；

SEQ ID NO：23(CNP-39)：

LLQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC；

SEQ ID NO：24(CNP-38)：

LQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC；

SEQ ID NO：25(CNP-37)：

QEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC；

SEQ ID NO：26 (CNP-37Q1pQ, wherein pQ=pyroglutamic acids)：

pQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC；

SEQ ID NO：27(G-CNP-37)：

GQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC；

SEQ ID NO：28(P-CNP-37)：

PQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC；

SEQ ID NO：29(M-CNP-37)：

MQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC；

SEQ ID NO：30(PG-CNP-37)：

PGQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC；

SEQ ID NO：31(MG-CNP-37)：

MGQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC；

SEQ ID NO：32(CNP-37M32N)：

QEHPNARKYKGANKKGLSKGCFGLKLDRIGSNSGLGC；

SEQ ID NO：33(G-CNP-37M32N)：

GQEHPNARKYKGANKKGLSKGCFGLKLDRIGSNSGLGC；

SEQ ID NO：34(G-CNP-37K14Q)：

GQEHPNARKYKGANQKGLSKGCFGLKLDRIGSMSGLGC；

SEQ ID NO：35(G-CNP-37K14P)：

GQEHPNARKYKGANPKGLSKGCFGLKLDRIGSMSGLGC；

SEQ ID NO：36 (G-CNP-37K14Q, Δs 15)：

GQEHPNARKYKGANQGLSKGCFGLKLDRIGSMSGLGC；

SEQ ID NO：37 (G-CNP-37K14Q, K15Q)：

GQEHPNARKYKGANQQGLSKGCFGLKLDRIGSMSGLGC；

SEQ ID NO：38(CNP-36)：

EHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC；

SEQ ID NO：39(CNP-35)：

HPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC；

SEQ ID NO：40(CNP-34)：

PNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC；

SEQ ID NO：41(CNP-33)：

NARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC；

SEQ ID NO：42(CNP-32)：

ARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC；

SEQ ID NO：43(CNP-31)：

RKYKGANKKGLSKGCFGLKLDRIGSMSGLGC；

SEQ ID NO：44(CNP-30)：

KYKGANKKGLSKGCFGLKLDRIGSMSGLGC；

SEQ ID NO：45(CNP-29)：

YKGANKKGLSKGCFGLKLDRIGSMSGLGC；

SEQ ID NO：46(CNP-28)：

KGANKKGLSKGCFGLKLDRIGSMSGLGC；

SEQ ID NO：47(GHKSEVAHRF-CNP-28)：

GHKSEVAHRFKGANKKGLSKGCFGLKLDRIGSMSGLGC；

SEQ ID NO：48(CNP-27)：

GANKKGLSKGCFGLKLDRIGSMSGLGC；

SEQ ID NO：49 (CNP-27K4Q, K5Q)：

GANQQGLSKGCFGLKLDRIGSMSGLGC；

SEQ ID NO：50 (CNP-27K4R, K5R)：

GANRRGLSKGCFGLKLDRIGSMSGLGC；

SEQ ID NO：51 (CNP-27K4P, K5R)：

GANPRGLSKGCFGLKLDRIGSMSGLGC；

SEQ ID NO：52 (CNP-27K4S, K5S)：

GANSSGLSKGCFGLKLDRIGSMSGLGC；

SEQ ID NO：53 (CNP-27K4P, K5R)：

GANGANPRGLSRGCFGLKLDRIGSMSGLGC；

SEQ ID NO：54 (CNP-27K4R, K5R, K9R)：

GANRRGLSRGCFGLKLDRIGSMSGLGC；

SEQ ID NO：55 (CNP-27K4R, K5R, K9R, M22N)：

GANRRGLSRGCFGLKLDRIGSNSGLGC；

SEQ ID NO：56 (P-CNP-27K4R, K5R, K9R)：

PGANRRGLSRGCFGLKLDRIGSMSGLGC；

SEQ ID NO：57 (M-CNP-27K4R, K5R, K9R)：

MGANRRGLSRGCFGLKLDRIGSMSGLGC；

SEQ ID NO：58 (HSA fragments-CNP-27)：

GHKSEVAHRFKGANKKGLSKGCFGLKLDRIGSMSGLG；

SEQ ID NO：59 (HSA fragments-CNP-27M22N)：

GHKSEVAHRFKGANKKGLSKGCFGLKLDRIGSNSGLGC；

SEQ ID NO：60 (M-HSA fragments-CNP-27)：

MGHKSEVAHRFKGANKKGLSKGCFGLKLDRIGSMSGLGC；

SEQ ID NO：61 (P-HSA fragments-CNP-27)：

PGHKSEVAHRFKGANKKGLSKGCFGLKLDRIGSMSGLGC；

SEQ ID NO：62(CNP-26)：

ANKKGLSKGCFGLKLDRIGSMSGLGC；

SEQ ID NO：63(CNP-25)：

NKKGLSKGCFGLKLDRIGSMSGLGC；

SEQ ID NO：64(CNP-24)：

KKGLSKGCFGLKLDRIGSMSGLGC；

SEQ ID NO：65(CNP-23)：

KGLSKGCFGLKLDRIGSMSGLGC；

SEQ ID NO：66(R-CNP-22)：

RGLSKGCFGLKLDRIGSMSGLGC；

SEQ ID NO：67(ER-CNP-22)：

ERGLSKGCFGLKLDRIGSMSGLGC；

SEQ ID NO：68(R-CNP-22K4R)：

RGLSRGCFGLKLDRIGSMSGLGC；

SEQ ID NO：69(ER-CNP-224KR)：

ERGLSRGCFGLKLDRIGSMSGLGC；

SEQ ID NO：70(RR-CNP-22)：

RRGLSRGCFGLKLDRIGSMSGLGC；

SEQ ID NO：71 (HRGP fragments-CNP-22)：

GHHSHEQHPHGANQQGLSKGCFGLKLDRIGSMSGLGC；

SEQ ID NO.72 (HRGP fragments-CNP-22)：

GAHHPHEHDTHGANQQGLSKGCFGLKLDRIGSMSGLGC；

SEQ ID NO：73 (HRGP fragments-CNP-22)：

GHHSHEQHPHGANPRGLSKGCFGLKLDRIGSMSGLGC；

SEQ ID NO：74(IgG_l(F_c) fragment-CNP-22)：

GQPREPQVYTLPPSGLSKGCFGLKLDRIGSMSGLGC；

SEQ ID NO：75 (HSA fragments-CNP-22)：

GQHKDDNPNLPRGANPRGLSKGCFGLKLDRIGSMSGLGC；

SEQ ID NO：76 (HSA fragments-CNP-22)：

GERAFKAWAVARLSQGLSKGCFGLKLDRIGSMSGLGC；

SEQ ID NO：77 (people's bone knits plain NPR C inhibitor fragments-CNP22)：

FGIPMDRIGRNPRGLSKGCFGLKLDRIGSMSGLGC；

SEQ ID NO：78 (FGF2 heparin binding domain fragments-CNP22)：

GKRTGQYKLGSKTGPGPKGLSKGCFGLKLDRIGSMSGLGC；

SEQ ID NO：79(IgG_l(F_e) fragment-CNP-22K4R)：

GQPREPQVYTGANQQGLSRGCFGLKLDRIGSMSGLGC；

SEQ ID NO：80 (HSA fragments-CNP-22K4R)：

GVPQVSTSTGANQQGLSRGCFGLKLDRIGSMSGLGC；

SEQ ID NO：81 (CH-296-GNP-22K4R)：

GQPSSSSQSTGANQQGLSRGCFGLKLDRIGSMSGLGC；

SEQ ID NO：82 (CH-296-CNP-22K4R)：

GQTHSSGTQSGANQQGLSRGCFGLKLDRIGSMSGLGC；

SEQ ID NO：83 (CH-296-CNP-22K4R)：

GSTGQWHSESGANQQGLSRGCFGLKLDRIGSMSGLGC；

SEQ ID NO：84 (zinc finger fragment-CNP-22K4R)：

GSSSSSSSSSGANQQGLSRGCFGLKLDRIGSMSGLGC；

SEQ ID NO：85(CNP-21)：

LSKGCFGLKLDRIGSMSGLGC；

SEQ ID NO：86(CNP-20)：

SKGCFGLKLDRIGSMSGLGC；

SEQ ID NO：87(CNP-19)：

KGCFGLKLDRIGSMSGLGC；

SEQ ID NO：88(CNP-18)：

GCFGLKLDRIGSMSGLGC；

SEQ ID NO：89(CNP-17)：

CFGLKLDRIGSMSGLGC；

SEQ ID NO：90 (BNP fragment-CNP-17-BNP fragments)：

SPKMVQGSGCFGLKLDRIGSMSGLGCKVLRRH；

SEQ ID NO：91(CNP-38L1G)：

GQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC；

SEQ ID NO：92(Ac-CNP-37；Wherein Ac=acetyl group)

Ac-QEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC；

It should be understood that SEQ ID NO：The equivalent of cysteine in 1 position 6 and 22 is in SEQ ID NOs：In 2 to 92 Connected also by disulphide bridges.

It is highly preferred that term " CNP " refers to SEQ ID：NOs 2、19、20、21、22、23、24、25、26、30、32、38、 39th, 40,41,42,43,91,92 sequence.Even further preferably, term " CNP " refers to SEQ ID：NOs 23、24、25、26、 38th, 39,91 and 92 sequence.In particularly preferred embodiments, term " CNP " refers to SEQ ID NO：24 sequence.

In a particularly preferred embodiment, term " CNP " refers to SEQ ID NO：23rd, 24,25 and 38 sequence, very To more preferably SEQ ID NO：24 and 25 sequence, most preferably SEQ ID NO：25 sequence.Also, it is preferred that embodiment In, term " CNP " is SEQ ID NO：24 sequence.

In another preferred embodiment, term " CNP " refers to SEQ ID NO：93 sequence

QEHPNARX₁YX₂GANX₃X₄GLSX₅GCFGLX₆LDRIGSMSGLGC,

Wherein X₁、X₂、X₃、X₄、X₅And X₆K, R, P, S and Q are independently selected from, on condition that at least X₁、X₂、X₃、X₄、X₅With X₆In one be selected from R, P, S and Q；Preferably X₁、X₂、X₃、X₄、X₅And X₆Selected from K and R, on condition that at least X₁、X₂、X₃、X₄、X₅ And X₆In one be R；

Even more preferably still refer to SEQ ID NO:94 sequence

QEHPNARKYKGANX₁X₂GLSX3GCFGLX₄LDRIGSMSGLGC,

Wherein X₁、X₂、X₃And X₄K, R, P, S and Q are independently selected from, on condition that at least X₁、X₂、X₃And X₄In one Selected from R, P, S and Q；Preferably X₁、X₂、X₃And X₄Selected from K and R, on condition that at least X₁、X₂、X₃And X₄In one be R；

And most preferably refer to SEQ ID NO:95 sequence

QEHPNARKYKGANX₁X₂GLSKGCFGLKLDRIGSMSGLGC,

Wherein X₁X₂Selected from KR, RK, KP, PK, SS, RS, SR, QK, QR, KQ, RQ, RR and QQ.

It should be appreciated that in all CNP sequences provided in this manual, SEQ ID NO:Half Guang of 1 position 6 and 22 The equivalent of propylhomoserin is in SEQ ID NO:Connected in 93 to 95 also by disulphide bridges.

It should be appreciated that present invention additionally comprises CNP variants, any of which are one or more until all easily by deamidation or de- The residue that acid amides sample reaction (for example, isomerization) influences can be reacted to any degree (up to by deamidation or deamidation sample 100% residue for converting/being converted) it is converted into other residues.In certain embodiments, the disclosure includes CNP variants, its In：

(1) any one or more are until whole asparagine (Asn/N) residues can be converted into asparagus fern by deamidation Propylhomoserin or aspartate, and/or different aspartic acid or different aspartate, be up to about 5%, 10%, 20%, 30%, 40%, 50%th, 60%, 70%, 80%, 90% or 100% residue for converting/being converted；Or

(2) any one or more are until whole glutamine (Gln/Q) residues can be converted into paddy ammonia by deamidation Acid or glutamate, and/or isoglutamic acid or isoglutamic acid salt, be up to about 5%, 10%, 20%, 30%, 40%, 50%, 60%th, 70%, 80%, 90% or 100% residue for converting/being converted；Or

(3) any one or more are until whole aspartic acids or aspartate (Asp/D) residue can be by de- Acid amides sample reaction (also referred to as isomerization) is converted into different aspartic acid or different aspartate, be up to about 5%, 10%, 20%, 30%th, 40%, 50%, 60%, 70%, 80%, 90% or 100% residue for converting/being converted；Or

(4) any one or more are until whole glutamic acid or glutamate (Glu/E) residue can pass through deamidation Sample reaction (also referred to as isomerization) is converted into isoglutamic acid or isoglutamic acid salt, be up to about 5%, 10%, 20%, 30%, 40%, 50%th, 60%, 70%, 80%, 90% or 100% residue for converting/being converted；

(5) N- terminal glutamins (if present) can be converted into pyroglutamic acid, be up to about 5%, 10%, 20%, 30%th, 40%, 50%, 60%, 70%, 80%, 90% or 100% conversion；Or

(5) combinations of the above.

As used herein, term " CNP polypeptide variants " refers to different from the more of same species from reference to CNP polypeptides Peptide.Preferably, such reference CNP peptide sequences are SEQ ID NO:1 sequence.Also, it is preferred that embodiment in, ginseng It is SEQ ID NO to examine CNP peptide sequences:24 sequence.Generally, difference is restricted, so as to reference to the amino acid sequence with variant It is similar on the whole to be listed in, and is identical in many regions.Preferably, CNP polypeptide variants and reference CNP polypeptides are excellent Select SEQ ID NO:1 CNP polypeptides are the identicals of at least 70%, 80%, 90% or 95%.Also, it is preferred that embodiment In, CNP polypeptide variants and the preferred SEQ ID NO of reference CNP polypeptides:24 CNP polypeptides be at least 70%, 80%, 90% or 95% identical.For the polypeptide with the amino acid sequence with inquiry (query) amino acid sequence at least 95% " identical ", meaning It is identical with search sequence to refer to the amino acid sequence of theme polypeptide, simply it is every can to include inquiry amino acid sequence for theme peptide sequence The change of up to 5 amino acid of 100 amino acid.Ammonia in reference amino acid sequence can occur for these changes of reference sequences Any position between base (N- ends) or carboxyl terminal (C- ends) position or these terminal positions, is respectively dispersed in reference In the continuous group of one or more between the residue of sequence or in reference sequences.Search sequence can be the whole of reference sequences Amino acid sequence or any specified segment as described herein.Preferably, search sequence is SEQ ID NO:1 sequence.Same In preferred embodiment, search sequence is SEQ ID NO:24 sequence.

Such CNP polypeptide variants can be naturally occurring variant, for example, by occupy on chromosome or organism to Determine the naturally occurring allele variant of one of CNP of several alternative forms of locus codings, or by from single first The isotype of the naturally occurring splice variant coding of level transcription.Or CNP polypeptide variants can be do not know it is naturally occurring simultaneously And the variant that can be prepared by induced-mutation technique known in the art.

It is known in the art, can be from the N- ends or C- terminal deletion one or more amino of biologically active peptide or protein Acid, without significantly losing biological function.This N- and/or C- terminal deletions are also included within term CNP polypeptide variants.

Those of ordinary skill in the art are also recognized that, thus it is possible to vary some amino acid sequences of CNP polypeptides, without notable shadow Ring the structure or function of peptide.Such mutant includes according to the missing of general rule known in the art selection, insertion, fallen Put, repeat and substitute, so as to be had little to no effect to activity.For example, Bowie et al. (1990), Science 247:1306- 1310 provide the guidance of the 49-Phe ,82-Ser,115-Arg,144-Met,145-Asn ,161-Arg,169-Met Human Connective tissue growth factor on how to carry out Phenotypic silence, and entire contents are incorporated herein by reference, Wherein author points out that research changes two main methods of the tolerance of amino acid sequence.

Term CNP polypeptides also include more by all CNP of CNP analogs, ortholog thing and/or Species homologues coding Peptide.As used herein, term " CNP analogs " refers to perform identical function in each organism but not shared from biological ancestors Ancestral structure different and incoherent biological CNP.Instead, similar CNP individually occurs, and then evolves as and holds The same or analogous function of row.In other words, similar CNP polypeptides are that have entirely different amino acid sequence but perform identical The bioactivity i.e. polypeptide of the growth of regulation chondrosin long slab cartilage cell, propagation and differentiation.

As used herein, term " CNP ortholog things " refers to the CNP in two kinds of different plant species (species), its sequence It is relative to each other by the common homologous CNP in progenitor species, but has entered to turn to different from each other.

As used herein, term " CNP homologues " refers to perform identical function in each organism and originating from biology The CNP of the different organisms for the ancestral structure that ancestors share.In other words, homology CNP polypeptides are that have closely similar amino The polypeptide of acid sequence (it performs identical bioactivity, that is, adjusts growth, propagation and the differentiation of chondrosin long slab cartilage cell). Preferably, CNP polypeptide homologs can be defined as and refer to the preferred SEQ ID NO of CNP polypeptides:1 CNP polypeptides are presented at least 40%th, the polypeptide of 50%, 60%, 70%, 80%, 90% or 95% homogeneity.Also, it is preferred that embodiment in, reference CNP polypeptides are SEQ ID NO:24 CNP polypeptides.

Therefore, can be for example according to the CNP polypeptides of the present invention：(i) wherein at least one amino acid residue guarded or The CNP polypeptides of the preferably conservative amino acid residue substitution of non-conservative amino acid residue, and the substituted amino acid residue It can be or can not be the residue by genetic code encoding；And/or (ii) wherein at least one amino acid residue includes substitution The CNP polypeptides of base；And/or (iii) wherein CNP polypeptides and another compound (such as the compound of the half-life period of increase polypeptide (such as polyethylene glycol)) fusion CNP polypeptides；And/or the CNP polypeptides of (iv) wherein other amino acid and CNP peptide fusions, Such as IgG Fc corresponding circle of sensation peptides or targeting sequencing or the sequence or precursor egg of secretion sequence or the polypeptide for purifying above-mentioned form Bai Xulie.

As used herein, term " CNP polypeptide fragments " refers to include the preferred SEQ IDNO of CNP polypeptides:The amino of 1 polypeptide Any peptide of the continuous span of a part for acid sequence.Also, it is preferred that embodiment in, term " CNP polypeptide fragments " refers to Include SEQ ID NO:Any peptide of the continuous span of a part for the amino acid sequence of 24 polypeptide.

More specifically, CNP polypeptide fragments include CNP polypeptides more preferably SEQ ID NO:At least six of 1 polypeptide, such as At least eight, at least ten or at least 17 continuous amino acid.It is also preferable that CNP polypeptide fragments include SEQ ID NO:24 At least six of CNP polypeptides, for example, at least 8, at least ten or at least 17 continuous amino acid.CNP polypeptide fragments can be another The outer subgenus for being described as the CNP polypeptides comprising at least six amino acid, wherein " at least 6 " are defined as 6 C with representing CNP polypeptides Any integer between the integer of end amino acid, the preferred SEQ ID NO of CNP polypeptides:1 polypeptide, or also, it is preferred that SEQ ID NO:24 polypeptide.Also include the species (species) that length as described above is at least the CNP polypeptide fragments of 6 amino acid, It is further designated with regard to their N- ends and C- terminal positions.Term " CNP polypeptide fragments " also includes as individual species Length as described above is at least all CNP polypeptide fragments of 6 amino acid, and it can be especially by N- ends and C- terminal positions Specify.That is, it is the continuous amino of at least six that length is can take up on any given amino acid sequence of CNP polypeptides The N- ends of the fragment of sour residue and each combination of C- terminal positions are included in the invention, the preferred SEQ of CNP polypeptides ID NO:1 CNP polypeptides, also, it is preferred that SEQ ID NO:24 CNP polypeptides.

Term " CNP " is also included with sequence as described above but with comprising amido link and the connection of non-acid amides (linkage) such as poly- (amino acid) conjugate of the main chain of ester connection, such as depsipeptides.Depsipeptides is the chain of amino acid residue, its Middle main chain includes acid amides (peptide) and ester bond.Therefore, as used herein, term " side chain " refers to the α-carbon for being connected to amino acid moiety Part, if amino acid moiety is connected by amine key in such as polypeptide, or refer to be connected to poly- (amino acid) conjugate Main chain on any part for including carbon atom, such as in the case of depsipeptides.Preferably, term " CNP " refers to have and led to The polypeptide for the main chain that superamide (peptide) key is formed.

Term CNP includes above-mentioned CNP variants, analog, ortholog thing, homologue, derivative and fragment, Suo Youcan That examines ad-hoc location in sequence quotes variant, analog, ortholog thing, homologue, derivative and the piece for also including CNP parts Equivalent site in section, even if not specifically mentioned.

As used herein, term " loop section " refers to positioned at two cysteine residues for forming intramolecular disulfide compound bridge Between or pass through the CNP medicines between the homology amino acid residue that chemical linker connects or partial continuous amino acid residue Extension.Preferably, loop section is located at and formed between two cysteine residues of intramolecular disulfide compound bridge.The two half Guang ammonia Acid corresponds to CNP-38 (SEQ ID NO:24) cysteine of 22 and 38 in sequence.Therefore, if CNP medicines or part Sequence with CNP-38, then amino acid 23 to 37 be located at the loop section.

Unrelated with the length of CNP parts, the sequence of wild type CNP loop section is FGLKLDRIGSMSGLG (SEQ ID NO:96)。

As described above, term " CNP " is related to CNP medicines or part with different number amino acid.People in the art Member understands, in the CNP medicines of different length or part, the position of equivalent amino acid is different, and those skilled in the art will be not difficult It is determined that form two cysteines of disulphide bridges or lead in CNP versions that are longer, shorter and/or otherwise modifying Cross their two homology amino acid residues that chemical linker (linker) is connected to each other.

Term CNP includes above-mentioned CNP variants, analog, ortholog thing, homologue, derivative and fragment, term " loop section " also includes SEQ ID NO:The variation of 96 sequence, analog, ortholog thing, homologue, derivative and Fragment.Therefore, in reference sequences ad-hoc location it is all quote also including the variants of CNP parts, analog, ortholog thing, Equivalent site in homologue, derivative and fragment, even if not specifically mentioned.

As used herein, term " pharmaceutical composition " refers to composition, and it includes one or more active components, such as medicine Thing or prodrug, the CNP prodrugs of the present invention are particularly herein；And optional one or more excipient；And directly or indirectly Solution caused by ground is planted into subassembly, complexing or aggregation by any two of composition or more or by one or more compositions From any product caused by caused or other types of reaction as one or more compositions or interaction.Therefore, this hair Bright pharmaceutical composition includes the one or more CNP prodrugs and optional pharmaceutically acceptable figuration by mixing the present invention Any combinations thing prepared by agent.

As used herein, term " fluid composition " refers to comprising water-soluble CNP prodrugs and one or more solvents for example The mixture of water.

Term " suspension composite " is related to mixture, and it includes water-insoluble CNP prodrugs, wherein for example carrier Z ' is water Gel, and one or more solvents, such as water.Due to insoluble polymer, polymeric prodrugs can not dissolve and make precursor Medicine is in graininess.

As used herein, term " dry composition " refers to the pharmaceutical composition provided in a dry form.Suitable drying side Method is spray drying and freezed, that is, is freeze-dried.The dry composition of this prodrug has according to karl Fischer (Karl Fischer what is) determined is up to 10%, is preferably smaller than 5%, more preferably less than 2% residual moisture content.Preferably, it is of the invention Pharmaceutical composition pass through lyophilization.

Term " medicine " used herein refers to be used to treat, cure, prevent or diagnose the illness or for otherwise Strengthen body or the material of mental health.If a kind of medicine is conjugated with another part (moiety), the source of obtained product It is referred to as " biologically-active moiety " from the part of medicine.

As used herein, term " prodrug " refer to by reversible prodrug connector part reversibly and be covalently attached to specially The biologically-active moiety of the protection group of door, described reversible prodrug connector part are comprising reversible with biologically-active moiety Couple the connector part of (linkage), and wherein special blocking group changes or eliminated undesirable in parent molecule Characteristic.This also includes desired property in increase medicine and suppresses undesirable property.Special nontoxic blocking group is referred to as " carrying Body ".Prodrug discharges reversible and covalently bound biologically-active moiety in the form of its relative medicine.In other words, prodrug is bag Conjugate containing biologically-active moiety by reversible prodrug connector some covalent and reversibly conjugated with carrier part, carry Body is covalent and reversible conjugated directly or by interval body (spacer) progress with reversible prodrug connector part.This conjugate The previously conjugated biologically-active moiety of release in the form of free drug.

" biodegradable connection " or " reversible connection " is (aqueous buffer solution, pH 7.4,37 in physiological conditions DEG C) under be not present degradable and water soluble (i.e. cleavable) connection in the case of enzyme, half-life period is 1 hour to 6 months, preferably 1 Hour to 4 months, even more preferably 1 hour to 3 months, even more preferably 1 hour to 2 months, even more preferably 1 hour to 1 Individual month.Therefore, stable connection is in physiological condition (pH7.4,37 DEG C of aqueous buffer solution) connection of the half-life more than 6 months Connect.

Therefore, " reversible prodrug connector part " is such part：It is covalently conjugated to biology by reversible connection Active part such as CNP and carrier part such as-Z or-Z' is also covalently conjugated to, wherein covalently sewing with the carrier part Close directly or by interval body portion such as-L²- realize.Preferably ,-Z or-Z' and-L²- between connection be stable connection.

As used herein, term " seamless prodrug connector " refers to reversible prodrug connector, and it is swum once cutting with it Medicine is discharged from form.As used herein, the medicine of term " free form " refers to that its is unmodified, pharmacological activity form Medicine.

As used herein, it is term " excipient " refers to apply together with therapeutic agent such as medicine or prodrug diluent, auxiliary Auxiliary agent or solvent.Such drug excipient can be sterile liquid, such as water and oil, including oil, animal, plant or synthesis Those of source, including but not limited to peanut oil, soybean oil, mineral oil, sesame oil etc..When orally administering pharmaceutical composition, Water is preferable excipient.When intravenously applying pharmaceutical composition, salt solution and D/W are preferable excipient.Salt The aqueous solution and D/W and glycerite are preferably used as the liquid excipient of Injectable solution.Suitable drug excipient Including starch, glucose, lactose, sucrose, mannitol, trehalose, gelatin, malt, rice, flour, chalk, silica gel, odium stearate, Glycerin monostearate, talcum, sodium chloride, dry skimmed milk, glycerine, propane diols, water, ethanol etc..If desired, pharmaceutical composition A small amount of wetting agent or emulsifying agent, pH buffer etc., such as acetate, succinate, tris, carbonate, phosphorus can also be contained Hydrochlorate, HEPES (4- (2- hydroxyethyls) -1- piperazine ethanesulfonic acids), MES (2- (N- morpholinoes) ethyl sulfonic acid) or can contain wash Wash agent such as tween, poloxamer, pool Lip river sand amine, CHAPS, Igepal or amino acid such as glycine, lysine or histidine. These pharmaceutical compositions can take the shape of solution, suspension, emulsion, tablet, pill, capsule, powder, extended release preparation etc. Formula.Pharmaceutical composition can be configured to suppository with traditional adhesive and excipient such as triglycerides.Oral formulations can include Standard excipients, such as the mannitol of pharmaceutical grade, lactose, starch, magnesium stearate, saccharin sodium, cellulose, magnesium carbonate etc..So Composition by medicine or biologically-active moiety containing therapeutically effective amount, and proper amount of excipient, to be carried for patient For appropriate administration form.Preparation should meet method of application.

As used herein, term " reagent " refers to comprising at least one functional group being used for another compound or medicine The compound of the functional group of reaction.It should be understood that the medicine comprising functional group's (such as primary amine or secondary amine or hydroxy functional group) is also Reagent.

As used herein, term " part (moiety) " refers to point for lacking one or more atoms compared with corresponding reagent A part (part) for son.For example, if the reagent of formula " H-X-H " is with another reagent reacting and as a part for reaction product (part), then the appropriate section (moiety) of reaction product has structure " H-X-" or "-X-", and each "-" represents to be connected to Another part (moiety).Therefore, biologically-active moiety discharges as medicine from prodrug.

It should be appreciated that armed with the sequence or chemical constitution of one group of atom, this group of atom be connected to two parts or Part (moiety) is interrupted, then the sequence or chemical constitution can be connected to two parts with any orientation, unless in addition Clearly state.For example, part "-C (O) N (R¹²)-", can be used as "-C (O) N (R¹²)-" or "-N (R¹²) C (O)-" be connected to Two parts or by partial interruption.Similarly, part

Can conductIt is connected to two parts or can be by partial interruption.

As used herein, term " functional group " refers to the atomic radical that can be reacted with other atomic radicals.Functional group wraps Include but be not limited to following group：Carboxylic acid (- (C=O) OH), primary or secondary amine (- NH₂,-NH -), maleimide, mercaptan (SH), sulphur Sour (- (O=S=O) OH), carbonic ester, carbamate (- O (C=O) N<), hydroxyl (- OH), aldehyde (- (C=O) H), ketone (- (C =O) -), (>N-N<), isocyanates, isothiocyanates, phosphoric acid (- O (P=O) OHOH), phosphonic acids (- O (P=O) OHH), halo Acetyl group, alkyl halide, acryloyl group, aryl fluoride, azanol, disulphide, sulfonamide, sulfuric acid, vinyl sulfone, vinyl ketone, again Azane, oxirane and aziridine.

It is corresponding present invention additionally comprises its in the case where the prodrug of the present invention includes one or more acid or basic groups Acceptable salt pharmaceutically or in toxicology, particularly its pharmaceutically available salt.Therefore, according to the invention, it is possible to use Prodrug of the invention comprising acidic-group, such as alkali metal salt, alkali salt or ammonium salt.These salt it is more accurate Example include sodium salt, sylvite, calcium salt, magnesium salts or with ammonia or organic amine such as ethamine, monoethanolamine, triethanolamine or amino acid Salt.The prodrug of the invention for the group that can be protonated comprising one or more basic groups may have and can be according to this hair It is bright to be used in the form of it is with inorganic or organic acid addition salts.The example of suitable acid include hydrogen chloride, hydrogen bromide, phosphoric acid, Sulfuric acid, nitric acid, methanesulfonic acid, p-methyl benzenesulfonic acid, naphthalenedisulfonic acid, oxalic acid, acetic acid, tartaric acid, lactic acid, salicylic acid, benzoic acid, first Acid, propionic acid, neopentanoic acid, diethyl acid acetic acid, malonic acid, butanedioic acid, pimelic acid, fumaric acid, maleic acid, malic acid, amino sulphur Acid, phenylpropionic acid, gluconic acid, ascorbic acid, isonicotinic acid, citric acid, adipic acid and well known by persons skilled in the art other Acid.For those skilled in the art, it is also known that have other methods that basic group is converted into cation, such as the alkyl of amine groups Change, obtain the appropriate counter ion counterionsl gegenions of positively charged ammonium group and salt.If the prodrug of the present invention is simultaneously comprising acid and alkalescence Group, in addition to mentioned salt form, present invention additionally comprises inner salt or glycine betaine (betaine) (amphion).Each salt can To be obtained by conventional method well known by persons skilled in the art, such as by the way that these prodrugs and organic or inorganic acid or alkali are existed Contacted in solvent or dispersant, or by carrying out anion exchange or cation exchange with other salt.Present invention additionally comprises this hair All salt of bright prodrug, due to low physiological compatibility, it is not directly applied for medicine, but available for the centre of such as chemical reaction Body or for preparing pharmaceutically acceptable salt.

Term " pharmaceutically acceptable " refers to the material to be damaged when being applied to patient, and preferably by managing What mechanism such as EMA (Europe) and/or FDA (US) and/or any other national authority were ratified, for animal, preferably use In people.

As used herein, the combination of term " about " and numerical value is used to indicate to add and subtract no more than the numerical value from numerical value 10% scope and including end points, the 8% of more preferably no more than described numerical value, even more preferably no more than described numerical value 5%, the 2% of most preferably not more than described numerical value.For example, phrase " the about 200 " scopes for being used to representing 200+/- 10% and including End points, i.e., 180 to 220 scope and including end points；Preferably 200+/- 8%, i.e., 184 to 216 scope and including end points；Very Be 200+/- 5% to more preferably scope, i.e., 190 to 210 scope and including end points；Most preferably 200+/- 2%, i.e., 196 to 204 scope and including end points.It is appreciated that with " the about 20% " percentage represented do not indicate that " 20%+/- 10% " i.e. from 10% to 30%, " about 20% " and refer to 18 to 22% and including end points, i.e., numerical value 20 plus and subtract 10%.

As used herein, term " polymer " " refer to include by it is linear, circular, branched, crosslinking or dendroid in a manner of or its Combination by the molecule that the constitutional repeating unit of chemical key connection is monomer, its can be synthesis or biological source or Both combinations.It should be appreciated that polymer can also include one or more of the other chemical group and/or part, such as one Or multiple functional groups.Preferably, soluble polymer has at least 0.5kDa molecular weight, for example, at least 1kDa molecular weight, The molecular weight of at least 2kDa molecular weight, at least 3kDa or at least 5kDa molecular weight.If polymer is solvable, its is excellent It is at most 1000kDa, at most such as 750kDa, at most such as 500kDa, such as at most 300kDa, for example at most that choosing, which has, 200kDa, such as at most 100kDa molecular weight.It should be appreciated that for insoluble polymer, such as hydrogel, it is impossible to be provided with The molecular weight ranges of meaning.

As used herein, (polymeric) of term " polymer " " means to include one or more polymer or polymer Partial reagent or part.Polymeric reagent or part optionally can also include one or more of the other part, its preferred choosing From：

●C_1-50Alkyl, C_2-50Alkenyl, C_2-50Alkynyl, C_3-10Cycloalkyl, 3- to 10- circle heterocycles base, 8- to 11- members miscellaneous two Ring group, phenyl, naphthyl, indenyl, indanyl and tetrahydro naphthyl；And

● selected from following connection (linkage)

Wherein

Dotted line instruction is connected to the remainder of the part or reagent, and

- R and-R^aIt is independently selected from-H, methyl, ethyl, propyl group, butyl, amyl group and hexyl.

It will be appreciated by those skilled in the art that the polymerizate obtained from polymerisation not necessarily all has identical molecular weight, and It is to show molecular weight distribution.Therefore, in molecular weight ranges used herein, molecular weight, polymer the scope of number of monomers and Number of monomers in polymer is the molecular weight of the average number of number average molecular and monomer, i.e. polymer or polymer moieties Arithmetic average and the number of monomers of polymer or polymer moieties arithmetic mean of instantaneous value.

Therefore, in the polymer moieties comprising " x " monomeric unit, monomer is corresponded to for any integer that " x " is provided Arithmetic average.The integer range being located therein for the arithmetic average of " x " given arbitrary integer scope offer monomer. As " arithmetic average of " x " given about x " integer representation monomer is located at x+/- 10%, preferably x+/- 8%, more preferably x +/- 5%, most preferably in the integer range of x+/- 2%.

As used herein, term " number-average molecular weight " refers to the general mathematical average value of the molecular weight of individual polymer.

The term " water solubility " related to carrier used herein refers to one for the CNP prodrugs of the present invention when the carrier During part, it is soluble in 1 liter of water, is formed uniform in the 20 DEG C of at least 1g CNP prodrugs comprising this water-solubility carrier Solution.Therefore, the term related to carrier " water-insoluble " refers to the part for the CNP prodrugs of the present invention when the carrier When, it is soluble in 1 liter of water, is formed uniformly molten in 20 DEG C of CNP prodrugs comprising this water insoluble carrier less than 1g Liquid.

As used herein, term " hydrogel " refers to the hydrophilic or amphipathic polymer net being made up of homopolymer or copolymer Network, its due to exist covalent chemical crosslinking but it is insoluble.The crosslinking provides network structure and physical integrity.

As used herein, term " thermal gels " refers under low temperature (low temperature range is about 0 DEG C to about 10 DEG C) about It is liquid or low viscosity solution that the shear rate of 0.1/ second, which has at 25 DEG C less than 500cps viscosity, but in higher temperature (higher temperature is about 30 DEG C to about 40 DEG C, e.g., from about 37 DEG C) has under the shear rate of about 0.1/ second at 25 DEG C to be less than The compound of 10000cps viscosity higher.

As used herein, the term " based on PEG " related to part or reagent represents that the part or reagent include PEG.It is preferably based on PEG part or reagent includes at least 10% (w/w) PEG, for example, at least 20% (w/w) PEG, such as At least 30% (w/w) PEG, for example, at least 40% (w/w) PEG, for example, at least 50% (w/w), for example, at least 60 (w/w) PEG, example Such as at least 70% (w/w) PEG, at least 80% (w/w) PEG, for example, at least 90% (w/w) PEG, for example, at least 95%.Based on PEG Part or reagent residuals weight percentage be preferably selected from lower part and connection other parts：

●C_1-50Alkyl, C_2-50Alkenyl, C_2-50Alkynyl, C_3-10Cycloalkyl, 3- to 10- circle heterocycles base, 8- to 11- members miscellaneous two Ring group, phenyl, naphthyl, indenyl, indanyl and tetrahydro naphthyl；With

● selected from following connection

Wherein

As used herein, institute is meaned to part or the related term of reagent " include at least X%PEG based on PEG's " State part or reagent includes at least X% (w/w) ethylene glycol unit (- CH₂CH₂O -), wherein ethylene glycol unit can be in part or examination (blockwise) blocked in agent, it is alternately arranged or can be with random distribution, all ethylene glycol lists of preferably described part or reagent Member is present in a block；The residuals weight percentage of part or reagent based on PEG is other parts, is preferably selected from following Part and connection：

● selected from following connection

Wherein

Correspondingly using term " comprise at least X% hyaluronic acids based on hyaluronic acid ".

Term " substituted " used herein refers to that molecule or partial one or more H atoms are referred to as " substituent " Homoatomic or atomic radical are not replaced.

Preferably, one or more further optionally substituents are independently from each other：Halogen ,-CN ,-COOR^x1、- OR^x1、-C(O)R^x1、-C(O)N(R^x1R^x1a)、-S(O)₂N(R^x1R^x1a)、-S(O)N(R^x1R^x1a)、-S(O)₂R^x1、-S(O)R^x1、-N (R^x1)S(O)₂N(R^x1aR^x1b)、-SR^x1、-N(R^x1R^x1a)、-NO₂、-OC(O)R^x1、-N(R^x1)C(O)R^x1a、-N(R^x1)S(O)₂R^x1a、-N(R^x1)S(O)R^x1a、-N(R^x1)C(O)OR^x1a、-N(R^x1)C(O)N(R^x1aR^x1b)、-OC(O)N(R^x1R^x1a)、-T⁰、C_1-50 Alkyl, C_2-50Alkenyl and C_2-50Alkynyl；Wherein-T⁰、C_1-50Alkyl, C_2-50Alkenyl and C_2-50Alkynyl is optionally by identical or different one Individual or multiple-R^x2Substitution, and wherein C_1-50Alkyl, C_2-50Alkenyl and C_2-50Alkynyl is optionally by one or more selected from following Group interrupts：

-T⁰-、-C(O)O-、-O-、-C(O)-、-C(O)N(R^x3)-、-S(O)₂N(R^x3)-、-S(O)N(R^x3)-、-S (O)₂-、-S(O)-、-N(R^x3)S(O)₂N(R^x3a)-、-S-、-N(R^x3)-、-OC(OR^x3)(R^x3a)-、-N(R^x3)C(O)N(R^x3a)- With-OC (O) N (R^x3)-；

-R^x1、-R^x1a、-R^x1bIt is independently selected from-H ,-T⁰、C_1-50Alkyl, C_2-50Alkenyl and C_2-50Alkynyl；Wherein-T⁰、 C_1-50Alkyl, C_2-50Alkenyl and C_2-50Alkynyl is optionally by identical or different one or more-R^x2Substitution, and wherein C_1-50Alkane Base, C_2-50Alkenyl and C_2-50Alkynyl is optionally interrupted by one or more selected from following group：-T⁰-、-C(O)O-、-O-、-C (O)-、-C(O)N(R^x3)-、-S(O)₂N(R^x3)-、-S(O)N(R^x3)-；-S(O)₂-、-S(O)-、-N(R^x3)S(O)₂N(R^x3a)-、- S-、-N(R^x3)-、-OC(OR^x3)(R^x3a)-、-N(R^x3)C(O)N(R^x3a)-and-OC (O) N (R^x3)-；

Each T⁰Independently selected from：Phenyl, naphthyl, indenyl, indanyl, tetrahydro naphthyl, C_3-10Cycloalkyl, 3- are miscellaneous to 10- members Ring group and 8- are to 11- 9-membered heterobicyclic bases；Wherein each T⁰Independently optionally by one or more identical or different-R^x2Substitution；

Respectively-R^x2Independently selected from：Halogen ,-CN, oxo (=O) ,-COOR^x4、-OR^x4、-C(O)R^x4、-C(O)N (R^x4R^x4a)、-S(O)₂N(R^x4R^x4a)、-S(O)N(R^x4R^x4a)、-S(O)₂R^x4、-S(O)R^x4、-N(R^x4)S(O)₂N(R^x4aR^x4b)、- SR^x4、-N(R^x4R^x4a)、-NO₂、-OC(O)R^x4、-N(R^x4)C(O)R^x4a、-N(R^x4)S(O)₂R^x4a、-N(R^x4)S(O)R^x4a、-N (R^x4)C(O)OR^x4a、-N(R^x4)C(O)N(R^x4aR^x4b)、-OC(O)N(R^x4R^x4a) and C_1-6Alkyl；Wherein C_1-6Alkyl is optionally by one Individual or multiple identical or different halogen substitutions；

Respectively-R^x3、-R^x3a、-R^x4、-R^x4a、-R^x4bIndependently selected from：- H and C_1-6Alkyl；Wherein C_1-6Alkyl is optionally by one Or multiple identical or different halogen substitutions.

It is highly preferred that one or more of further optionally substituents be independently selected from halogen ,-CN ,- COOR^x1、-OR^x1、-C(O)R^x1、-C(O)N(R^x1R^x1a)、-S(O)₂N(R^x1R^x1a),-S(O)N(R^x1R^x1a)、-S(O)₂R^x1、-S (O)R^x1、-N(R^x1)S(O)₂N(R^x1aR^x1b)、-SR^x1、-N(R^x1R^x1a)、-NO₂、-OC(O)R^x1、-N(R^x1)C(O)R^x1a、-N (R^x1)S(O)₂R^x1a、-N(R^x1)S(O)R^x1a、-N(R^x1)C(O)OR^x1a、-N(R^x1)C(O)N(R^x1aR^x1b)、-OC(O)N (R^x1R^x1a)、-T⁰、C_1-10Alkyl, C_2-10Alkenyl and C_2-10Alkynyl；Wherein-T⁰、C_1-10Alkyl, C_2-10Alkenyl and C_2-10Alkynyl is optional By identical or different one or more-R^x2Substitution, and wherein C_1-10Alkyl, C_2-10Alkenyl and C_2-10Alkynyl is optionally by one It is or multiple selected from following group interruption：-T⁰-、-C(O)O-、-O-、-C(O)-、-C(O)N(R^x3)-、-S(O)₂N(R^x3)-、-S (O)N(R^x3)-、-S(O)₂-、-S(O)-、-N(R^x3)S(O)₂N(R^x3a)-、-S-、-N(R^x3)-、-OC(OR^x3)(R^x3a)-、-N (R^x3)C(O)N(R^x3a)-and-OC (O) N (R^x3)-；

Respectively-R^x1、-R^x1a、-R^x1b、-R^x3、-R^x3aIndependently selected from：- H, halogen, C_1-6Alkyl, C_2-6Alkenyl and C_2-6Alkynyl；

Respectively-R^x4、-R^x4a、-R^x4bIndependently selected from：- H, halogen, C_1-6Alkyl, C_2-6Alkenyl and C_2-6Alkynyl；

Even further preferably, one or more of further optionally substituents be independently selected from halogen ,-CN ,- COOR^x1、-OR^x1、-C(O)R^x1、-C(O)N(R^x1R^x1a)、-S(O)₂N(R^x1R^x1a),-S(O)N(R^x1R^x1a)、-S(O)₂R^x1、-S (O)R^x1、-N(R^x1)S(O)₂N(R^x1aR^x1b)、-SR^x1、-N(R^x1R^x1a)、-NO₂、-OC(O)R^x1、-N(R^x1)C(O)R^x1a、-N (R^x1)S(O)₂R^x1a、-N(R^x1)S(O)R^x1a、-N(R^x1)C(O)OR^x1a、-N(R^x1)C(O)N(R^x1aR^x1b)、-OC(O)N (R^x1R^x1a)、-T⁰、C_1-6Alkyl, C_2-6Alkenyl and C_2-6Alkynyl；Wherein-T⁰、C_1-6Alkyl, C_2-6Alkenyl and C_2-6Alkynyl is optionally by phase Same or different one or more-R^x2Substitution, and wherein C_1-6Alkyl, C_2-6Alkenyl and C_2-6Alkynyl is optionally one or more Interrupted selected from following group：-T⁰-、-C(O)O-、-O-、-C(O)-、-C(O)N(R^x3)-、-S(O)₂N(R^x3)-、-S(O)N (R^x3)-、-S(O)₂-、-S(O)-、-N(R^x3)S(O)₂N(R^x3a)-、-S-、-N(R^x3)-、-OC(OR^x3)(R^x3a)-、-N(R^x3)C (O)N(R^x3a)-and-OC (O) N (R^x3)-；

Respectively-R^x1、-R^x1a、-R^x1b、-R^x2、-R^x3、-R^x3aIndependently selected from：- H, halogen, C_1-6Alkyl, C_2-6Alkenyl and C_2-6 Alkynyl；

Each T⁰Independently selected from：Phenyl, naphthyl, indenyl, indanyl, tetrahydro naphthyl, C_3-10Cycloalkyl, 3- are miscellaneous to 10- members Ring group and 8- are to 11- 9-membered heterobicyclic bases；Wherein each T⁰Independently optionally by one or more identical or different-R^x2Substitution.

Preferably, most 6-H atoms of the molecule optionally substituted are independently substituted base and substituted, such as 5-H atom Base is independently substituted to substitute, 4-H atom is independently substituted base and substituted, and 3-H atom is independently substituted base and substituted, and 2 Individual-H atom is independently substituted base and substituted, or 1-H atom is substituted base and substituted.

Term " interruption " refers between part (moiety) is inserted in into two carbon atoms, or if insertion positioned at part One end, then be inserted between carbon atom or hetero atom and hydrogen atom, be preferably inserted between carbon atom and hydrogen atom.

As used herein, term " C alone or in combination_1-4Alkyl " represents the straight or branched with 1 to 4 carbon atom Moieties.If present in molecular end, then straight or branched C_1-4The example of alkyl is methyl, ethyl, n-propyl, isopropyl Base, normal-butyl, isobutyl group, sec-butyl and the tert-butyl group.When two parts of molecule are by C_1-4When alkyl connects, these C_1-4Alkyl Example is-CH₂-、-CH₂-CH₂-、-CH(CH₃)-、-CH₂-CH₂-CH₂-、-CH(C₂H₅)-、-C(CH₃)₂-。C_1-4Alkyl carbon it is every Individual hydrogen can be substituted optionally by substituent as defined above.Optionally, C_1-4Alkyl can be by one or more defined below Partial interruption.

As used herein, term " C alone or in combination_1-6Alkyl " represents the straight or branched with 1 to 6 carbon atom Moieties.If present in molecular end, then straight chain and side chain C_1-6The example of alkyl is methyl, ethyl, n-propyl, isopropyl Base, normal-butyl, isobutyl group, sec-butyl, the tert-butyl group, n-pentyl, 2- methyl butyls, 2,2- dimethyl propyls, n-hexyl, 2- methyl Amyl group, 3- methyl amyls, 2,2- dimethylbutyls, 2,3- dimethylbutyls and 3,3- dimethyl propyls.When two portions of molecule Divide by C_1-6When alkyl connects, these C_1-6The example of alkyl is-CH₂-、-CH₂-CH₂-、-CH(CH₃)-、-CH₂-CH₂-CH₂-、- CH(C₂H₅)-and-C (CH₃)₂-。C_1-6Each hydrogen of carbon can be substituted optionally by substituent as defined above.Optionally, C_1-6 Alkyl can be by one or more partial interruptions defined below.

Therefore, " C_1-10Alkyl ", " C_1-20Alkyl " or " C_1-50Alkyl " represents there is 1 to 10,1 to 20 or 1 to 50 respectively The alkyl chain of carbon atom, wherein C_1-10、C_1-20Or C_1-50Each hydrogen atom of carbon can be replaced optionally by substituent as defined above Generation.Optionally, C_1-10Or C_1-50Alkyl can be by one or more partial interruptions defined below.

As used herein, term " C alone or in combination_2-6Alkenyl " is represented with 2 to 6 carbon atoms, comprising at least one The straight or branched hydrocarbon part of carbon-to-carbon double bond.If present in the end of molecule, then example is-CH=CH₂,-CH=CH- CH₃、-CH₂- CH=CH₂,-CH=CHCH₂-CH₃With-CH=CH-CH=CH₂.When two parts of molecule are by C_2-6Alkenyl connects When, such C_2-6The example of alkenyl is-CH=CH-.C_2-6Each hydrogen atom of alkenyl part can be optionally as defined above Substituent substitute.Optionally, C_2-6Alkenyl can be by one or more partial interruptions defined below.

Therefore, term " C alone or in combination_2-10Alkenyl ", " C_2-20Alkenyl " or " C_2-50Alkenyl " refers to there is 2 to 10,2 To 20 or 2 to 50 carbon atoms, the straight or branched hydrocarbon part comprising at least one carbon-to-carbon double bond.C_2-10Alkenyl, C_2-20Alkenyl Or C_2-50Each hydrogen atom of alkenyl can be substituted optionally by substituent as defined above.Optionally, C_2-10Alkenyl, C_2-20Alkene Base or C_2-50Alkenyl can be by one or more partial interruptions defined below.

As used herein, term " C alone or in combination_2-6Alkynyl " refers to 2 to 6 carbon atoms, comprising at least one The straight or branched hydrocarbon part of the key of carbon-to-carbon three.If present in the end of molecule, then example is-C ≡ CH ,-CH₂-C≡CH、 CH₂-CH₂- C ≡ CH and CH₂-C≡C-CH₃.When two parts of molecule are connected by alkynyl, then example is, for example ,-C ≡ C-. C_2-6Each hydrogen atom of alkynyl can be substituted optionally by substituent as defined above.It may be optionally present one or more Double bond.Optionally, C_2-6Alkynyl can be by one or more partial interruptions defined below.

Therefore, as used herein, term " C alone or in combination_2-10Alkynyl ", " C_2-20Alkynyl " and " C_2-50Alkynyl " represents There is 2 to 10,2 to 20 or 2 to 50 carbon atoms, the straight or branched hydrocarbon portion for including at least one key of carbon-to-carbon three respectively Point.C_2-10Alkynyl, C_2-20Alkynyl or C_2-50Each hydrogen atom of alkynyl can be substituted optionally by substituent as defined above.Appoint Selection of land, there may be one or more double bonds.Optionally, C_2-10Alkynyl, C_2-20Alkynyl or C_2-50Alkynyl can be by defined below One or more partial interruptions.

As described above, C_1-4Alkyl, C_1-6Alkyl, C_1-10Alkyl, C_1-20Alkyl, C_1-50Alkyl, C_2-6Alkenyl, C_2-10Alkenyl, C_2-20Alkenyl, C_2-50Alkenyl, C_2-6Alkynyl, C_2-10Alkynyl, C_2-20Alkenyl or C_2-50Alkynyl is optionally by one or more parts Disconnected, the part is preferably selected from：

Wherein

Dotted line instruction is connected to the remainder (remainder) of part or reagent；And

As used herein, term " C_3-10Cycloalkyl " refers to the cyclic alkyl chain with 3 to 10 carbon atoms, and it can be Saturation is undersaturated, such as cyclopropyl, cyclobutyl, cyclopenta, cyclohexyl, cyclohexenyl group, suberyl, cyclooctyl, cyclononyl Or cyclodecyl.C_3-10Each hydrogen atom of cycloalkyl carbon can be substituted by substituent as defined above.Term " C_3-10Cycloalkyl " is also Two rings such as norbornane or ENB including bridging.

Term " 8- to the more ring groups of 30- member carbon " or " 8- is polycyclic to 30- member carbon " refer to two with 8 to 30 annular atoms Or more ring loop section, two of which adjacent ring shares at least one annular atom, and can contain at most maximum quantity Double bond (aromatic series or non-aromatic ring, it is fully saturated, fractional saturation or undersaturated).Preferably, 8- to 30- The more ring groups of carbon of member refer to the loop section of 2,3,4 or 5 rings, the loop section of more preferably 2,3 or 4 rings.

Term " 3- to 10- circle heterocycles base " used herein or " 3- to 10- circle heterocycles " refer to there is 3,4,5,6,7,8,9 Or the ring of 10 annular atoms, (aromatic series or non-aromatic ring, it is fully saturated to its double bond containing at most maximum quantity , fractional saturation or it is undersaturated), wherein at least one annular atom until 4 annular atoms be chosen bin cure (including-S (O)-,- S(O)₂-), the hetero atom of oxygen and nitrogen (including=N (O) -) substitute, and wherein described ring passes through carbon or nitrogen-atoms and molecule Remainder connects.3- to the example of 10- circle heterocycles include but is not limited to aziridine, oxirane, thiirane, azirine, Oxireme, thiirene (thiirene), azetidine, oxetanes, Thietane (thietane), furan Mutter, thiophene, pyrroles, pyrrolin, imidazoles, imidazoline, pyrazoles, pyrazoline,Azoles,It is oxazoline, differentIt is azoles, differentOxazoline, thiophene Azoles, thiazoline, isothiazole, isothiazoline, thiadiazoles, Thiadiazoline, tetrahydrofuran, thiophane, pyrrolidines, imidazolidine, pyrrole Oxazolidine,It is oxazolidine, differentOxazolidine, thiazolidine, isothiazolidine, thiadiazolidine, sulfolane, pyrans, dihydropyran, oxinane, Imidazolidine, pyridine, pyridazine, pyrazine, pyrimidine, piperazine, piperidines, morpholine, tetrazolium, triazole, triazolidine, four oxazolidines, diaza cycloheptyl Alkane, azepineAnd homopiperazine.3- to 10- circle heterocycles base or 3- to each hydrogen atom of 10- circle heterocycles groups can be by following fixed The substituent of justice substitutes.

Term " 8- to 11- 9-membered heterobicyclics base " used herein or " 8- to 11- 9-membered heterobicyclics " refer to there is 8 to 11 rings The heterocyclic moiety of two rings of atom, wherein at least one annular atom is shared by two rings and it can contain at most maximum number The double bond (aromatic series or non-aromatic ring, it is fully saturated, fractional saturation or undersaturated) of amount, wherein at least one Annular atom until 6 annular atoms be chosen bin cure (including-S (O)-,-S (O)₂-), the hetero atom of oxygen and nitrogen (including=N (O) -) replaces Generation, and its middle ring is connected to the remainder of molecule by carbon or nitrogen-atoms.The example of 8- to 11- 9-membered heterobicyclics be indoles, Indoline, benzofuran, benzothiophene, benzoAzoles, benzisoxaAzoles, benzothiazole, benzisothiazole, benzimidazoles, benzene And imidazoline, quinoline, quinazoline, dihydroquinazoline, quinoline, EEDQ, tetrahydroquinoline, decahydroquinoline, isoquinolin, decahydro are different Quinoline, tetrahydroisoquinoline, dihydro-isoquinoline, benzo-azaPurine and pteridine.Term 8- is to 11- 9-membered heterobicyclics also including two The spiro connection structure of individual ring such as 1,4- dioxas -8- azaspiros [4.5] decane or bridged heterocyclic such as 8- aza-bicyclos [3.2.1] Octane.The substituent that each hydrogen atom of 8- to 11- 9-membered heterobicyclics base or 8- to 11- 9-membered heterobicyclic carbon can be defined as follows replaces Generation.

Similarly, term " 8- to 30- member miscellaneous more ring groups " or " 8 to 30 yuan miscellaneous polycyclic " refer to have 8 to 30 annular atoms, The heterocyclic moiety of preferably 3,4 or 5 rings of more than two ring, the adjacent ring of two of which share at least one annular atom, and And the double bond of at most maximum quantity can be contained (aromatic series or non-aromatic ring, it is fully saturated, fractional saturation or not Saturation), wherein at least one annular atom until 10 annular atoms be chosen bin cure (including-S (O)-,-S (O)₂-), oxygen and nitrogen The hetero atom of (including=N (O) -) is substituted, and wherein described ring is connected by the remainder of carbon or nitrogen-atoms and molecule.

It should be appreciated that and structure divisionRelated phrase " R^x/R^yThe shape pair together with the atom that they are connected Into C_3-10Cycloalkyl or 3- are to 10- circle heterocycles base " mean that Rx and Ry forms following structure：

Wherein R is C_3-10Cycloalkyl or 3- are to 10- circle heterocycles bases.

It should be appreciated that and structure divisionRelated phrase " R^x/R^yPair atom one being connected with them Rise and form ring A " and mean R^xAnd R^yForm following structure：

As used herein, term " end alkynes " means

Part.

As used herein, " halogen " means fluorine, chlorine, bromine or iodine.Generally preferably, halogen is fluorine or chlorine.

Generally, term " comprising " or "comprising" are also contemplated by " Consists of ".

Preferably ,-D has SEQ ID NO:24、SEQ ID NO:25 or SEQ ID NO:30 sequence, even more preferably SEQ ID NO:24 and SEQ ID NO:25 sequence.

In one embodiment ,-D has SEQ ID NO:25 sequence.

In another embodiment ,-D has SEQ ID NO:30 sequence.

In a preferred embodiment ,-D has SEQ ID NO:24 sequence.

Part-the L¹- it is reversible prodrug connector, medicine is that CNP is released from its free form, i.e., it is Seamless prodrug connector.Suitable prodrug connector is known in the art, such as is disclosed in WO 2005/099768 A2, WO Reversible prodrug connector parts of 2006/136586 A2, the WO 2011/089216 in the A1 and A1 of WO 2013/024053, It is incorporated herein by reference.

In another embodiment ,-L¹- it is 2011/089214 A1, the WO 2011/ of A1, WO of WO 2011/012722 Reversible prodrug connectors of 089215 A1, the WO 2013/024052 described in the A1 and A1 of WO 2013/160340, it passes through It is incorporated herein by reference.

Partly-L¹--D can be connected to by any kind of connection (linkage), on condition that it is reversible.It is preferred that Ground ,-L¹- pass through the connection selected from acid amides, ester, carbamate, acetal, aminal, imines, oxime, hydrazone, disulphide and acylguanidines It is connected in succession on-D.Even further preferably ,-L¹- be connected to by the connection selected from acid amides, ester, carbamate and acylguanidines- D。

In a preferred embodiment, part-L¹- be connected by acid amides connection with-D.It should be appreciated that acid amides connection is logical It is often irreversible, but in the present invention, included in-L¹- in adjacent group cause acid amides connection it is reversible.

Particularly preferred part-L¹- be disclosed in the A2 of WO 2009/095479.Therefore, in a preferred embodiment In, part-L¹- there is formula (II) structure：

Wherein dotted line instruction is by forming amido link connection most on the-D of CNP parts nitrogen；

- X- is-C (R⁴R^4a)-；-N(R⁴)-；-O-；-C(R⁴R^4a)-C(R⁵R^5a)-；-C(R⁵R^5a)-C(R⁴R^4a)-；-C (R⁴R^4a)-N(R⁶)-；-N(R⁶)-C(R⁴R^4a)-；-C(R⁴R^4a)-O-；-O-C(R⁴R^4a)-；Or-C (R⁷R^7a)-；

X¹It is C；Or S (O)；

-X²- it is-C (R⁸R^8a)-；Or-C (R⁸R^8a)-C(R⁹R^9a)-；

=X³It is=O；=S；Or=N-CN；

-R¹、-R^1a、-R²、-R^2a、-R⁴、-R^4a、-R⁵、-R^5a、-R⁶、-R⁸、-R^8a、-R⁹、-R^9aIndependently selected from-H；And C_1-6 Alkyl；

-R³、-R^3aIndependently selected from-H；And C_1-6Alkyl, if on condition that-R³、-R^3aOne or both of be not-H, then it Pass through SP³The carbon atom of-hydridization is connected on the N that they are connected；

-R⁷It is-N (R¹⁰R^10a)；Or-NR¹⁰- (C=O)-R¹¹；

-R^7a、-R¹⁰、-R^10a、-R¹¹It is-H independently of each other；Or C_1-6Alkyl；

Optionally, one or more-R^1a/-R^4a、-R^1a/-R^5a、-R^1a/-R^7a、-R^4a/-R^5a、-R^8a/-R^9aTo forming chemistry Key；

Optionally, one or more-R¹/-R^1a、-R²/-R^2a、-R⁴/-R^4a、-R⁵/-R^5a、-R⁸/-R^8a、-R⁹/-R^9aPair with The atom that they are connected forms C together_3-10Cycloalkyl；Or 3- is to 10- circle heterocycles bases；

Optionally, one or more-R¹/-R⁴、-R¹/-R⁵、-R¹/-R⁶、-R¹/-R^7a、-R⁴/-R⁵、-R⁴/-R⁶、-R⁸/- R⁹、-R²/-R³Pair ring A is formed together with the atom that they are connected；

Optionally, R³/R^3a3- is formed together with the nitrogen-atoms connected with them to 10- circle heterocycles；

A is selected from phenyl；Naphthyl；Indenyl；Indanyl；Tetrahydro naphthyl；C_3-10Cycloalkyl；3- is to 10- circle heterocycles bases；And 8- To 11- 9-membered heterobicyclic bases；And

Wherein-L¹- by-L²- Z or-L²- Z ' substitutes, and wherein-L¹- be optionally further substituted, on condition that formula (II) In with asterisk mark hydrogen not by-L²- Z or-L²- Z ' or substituent are replaced；

Wherein

-L²- it is single chemical bond or interval body；

- Z is water-solubility carrier；And

- Z ' is water insoluble carrier.

Preferably ,-the L of formula (II)¹- by a part-L²- Z or-L²- Z ' substitutes.

In one embodiment ,-the L of formula (II)¹- be not further substituted.

If it should be understood that-the R of formula (II)³/-R^3a3- is formed together with the nitrogen-atoms connected with them to 10- circle heterocycles, The atom for being only wherein connected directly to nitrogen is SP³Such 3- can be formed during the carbon atom of-hydridization to 10- circle heterocycles.In other words, By-R³/-R^3aThe such 3- formed together with the nitrogen-atoms connected with them has following structure to 10- circle heterocycles：

Wherein

Dotted line instruction is connected to-L¹- remainder (rest)；

The ring includes 3 to 10 atoms, including at least one nitrogen；And

R^#And R^##Represent SP³The carbon atom of-hydridization.

It is also understood that 3- can be further substituted to 10- circle heterocycles.

By formula (II)-R³/-R^3aThe suitable 3- formed together with the nitrogen-atoms connected with them is to 10- circle heterocycles Exemplary is as follows:

Wherein

Dotted line indicates to be connected to the remainder of molecule；And

- R is selected from-H and C_1-6Alkyl.

- the L of formula (II)¹- can optionally be further substituted.In general, any substituent can be used, as long as splitting Solution principle is not affected, i.e., the hydrogen marked with asterisk in formula (II) is not replaced and the part of formula (II)Nitrogen Remain the part (part) of primary, secondary or tertiary amine, i.e.-R³With-R^3aIt is independently of each other-H or passes through SP³The carbon atom of-hydridization It is connected to-N<.

In one embodiment ,-the R of formula (II)¹Or-R^1aBy-L²- Z or-L²- Z ' substitutes.In another embodiment In ,-the R of formula (II)²Or-R^2aBy-L²- Z or-L²- Z ' substitutes.In another embodiment ,-the R of formula (II)³Or-R^3aBy- L²- Z or-L²- Z ' substitutes.In another embodiment ,-the R of formula (II)⁴By-L²- Z or-L²- Z ' substitutes.In another implementation In scheme ,-the R of formula (II)⁵Or-R^5aBy-L²- Z or-L²- Z ' substitutes.In another embodiment ,-the R of formula (II)⁶By- L²- Z or-L²- Z ' substitutes.In another embodiment ,-the R of formula (II)⁷Or-R^7aBy-L²- Z or-L²- Z ' substitutes.Another In individual embodiment ,-the R of formula (II)⁸Or-R^8aBy-L²- Z or-L²- Z ' substitutes.In another embodiment, formula (II)- R⁹Or-R^9aBy-L²- Z or-L²- Z ' substitutes.

Most preferably ,-the R of formula (II)⁴By-L²- Z or-L²- Z ' substitutes.

Preferably ,-the X- of formula (II) is-C (R⁴R^4a)-or-N (R⁴)-.Most preferably ,-the X- of formula (II) is-C (R⁴R^4a)-。

Preferably, the X of formula (II)¹It is C.

Preferably, formula (II)=X³It is=O.

Preferably ,-the X of formula (II)²- it is-C (R⁸R^8a)-。

Preferably ,-the R of formula (II)⁸With-R^8aIndependently selected from-H, methyl and ethyl.More preferably-the R of formula (II)⁸With- R^8aIn it is at least one be-H.Even further preferably ,-the R of formula (II)⁸With-R^8aAll it is-H.

Preferably ,-the R of formula (II)¹With-R^1aIndependently selected from-H, methyl and ethyl.More preferably ,-the R of formula (II)¹ With-R^1aIn it is at least one be-H.Even further preferably ,-the R of formula (II)¹With-R^1aAll it is-H.

Preferably ,-the R of formula (II)²With-R^2aIndependently selected from-H, methyl and ethyl.It is highly preferred that-the R of formula (II)² With-R^2aIn it is at least one be-H.Even further preferably ,-the R of formula (II)²With-R^2aAll it is H.

Preferably ,-the R of formula (II)³With-R^3aIndependently selected from-H, methyl, ethyl, propyl group and butyl.Even more preferably The ground ,-R of formula (II)³With-R^3aIn it is at least one be methyl.In a same preferred embodiment ,-the R of formula (II)³With- R^3aAll it is-H.It is another also, it is preferred that embodiment in ,-the R of formula (II)³With-R^3aAll it is methyl.

Preferably ,-the R of formula (II)³It is-H, and-the R of formula (II)^3aIt is methyl.

Preferably ,-the R of formula (II)⁴With-R^4aIndependently selected from-H, methyl and ethyl.It is highly preferred that-the R of formula (II)⁴ With-R^4aIn it is at least one be-H.Even further preferably ,-the R of formula (II)⁴With-R^4aAll it is-H.

Preferably, part-L¹- there is formula (IIa) structure:

Wherein on the nitrogen for-D that dotted line instruction connects most CNP parts (moiety) by forming amido link；

-R¹、-R^1a、-R²、-R^2a、-R³、-R^3a、-R⁴、-R^4aWith-X²- used as defined in formula (II)；And

Wherein-L¹- by-L²- Z or-L²- Z ' substitutes and wherein-L¹- be optionally further substituted, on condition that formula (IIa) The middle hydrogen marked with asterisk is not by-L²- Z or-L²- Z ' or substituent are replaced.

Preferably ,-the L of formula (IIa)¹- by a part-L²- Z or-L²- Z ' substitutes.

Preferably, the part-L of formula (IIa)¹- be not further substituted.

Preferably ,-the R of formula (IIa)¹With-R^1aIndependently selected from-H, methyl and ethyl.It is highly preferred that-the R of formula (IIa)¹ With-R^1aIn it is at least one be-H.Even further preferably ,-the R of formula (IIa)¹With-R^1aAll it is-H.

Preferably ,-the R of formula (IIa)⁴With-R^4aIndependently selected from-H, methyl and ethyl.It is highly preferred that-the R of formula (IIa)⁴ With-R^4aIn it is at least one be-H.Even further preferably ,-the R of formula (IIa)⁴With-R^4aAll it is-H.

Preferably ,-the X of formula (IIa)²- it is-C (R⁸R^8a)-。

Preferably ,-the R of formula (IIa)⁸With-R^8aIndependently selected from-H, methyl and ethyl.It is highly preferred that-the R of formula (IIa)⁸ With-R^8aIn it is at least one be-H.Even further preferably ,-the R of formula (IIa)⁸With-R^8aAll it is-H.

Preferably ,-the R of formula (IIa)²With-R^2aIndependently selected from-H, methyl and ethyl.More preferably ,-the R of formula (IIa)² With-R^2aIn it is at least one be-H.Even further preferably ,-the R of formula (IIa)²With-R^2aAll it is H.

Preferably ,-the R of formula (IIa)³With-R^3aIndependently selected from-H, methyl, ethyl, propyl group and butyl.Even more preferably The ground ,-R of formula (IIa)³With-R^3aIn it is at least one be methyl.In a same preferred embodiment ,-the R of formula (IIa)³ With-R^3aAll it is-H.It is another also, it is preferred that embodiment in ,-the R of formula (IIa)³With-R^3aAll it is methyl.

Preferably ,-the R of formula (IIa)³It is-H, and-the R of formula (IIa)^3aIt is methyl.

Preferably, part-L¹- there is formula (IIb) structure:

-R²、-R^2a、-R³、-R^3aWith-X²- used as defined in formula (II)；And

Wherein-L¹- by-L²- Z or-L²- Z ' substitutes and wherein-L¹- be optionally further substituted,

On condition that the hydrogen of the asterisk mark in formula (IIb) is not by-L²- Z or-L²- Z ' or substituent are replaced.

Preferably ,-the L of formula (IIb)¹- by a part-L²- Z or-L²- Z ' substitutes.

Preferably, the part-L of formula (IIb)¹- be not further substituted.

Preferably ,-the X of formula (IIb)²- it is-C (R⁸R^8a)-。

Preferably ,-the R of formula (IIb)⁸With-R^8aIndependently selected from-H, methyl and ethyl.It is highly preferred that-the R of formula (IIb)⁸ With-R^8aIn it is at least one be-H.Even further preferably ,-the R of formula (IIb)⁸With-R^8aAll it is-H.

Preferably ,-the R of formula (IIb)²With-R^2aIndependently selected from-H, methyl and ethyl.More preferably ,-the R of formula (IIb)² With-R^2aIn it is at least one be-H.Even further preferably ,-the R of formula (IIb)²With-R^2aAll it is H.

Preferably ,-the R of formula (IIb)³With-R^3aIndependently selected from-H, methyl, ethyl, propyl group and butyl.Even more preferably The ground ,-R of formula (IIb)³With-R^3aIn it is at least one be methyl.In a same preferred embodiment ,-the R of formula (IIb)³ With-R^3aAll it is-H.It is another also, it is preferred that embodiment in ,-the R of formula (IIb)³With-R^3aAll it is methyl.

Most preferably ,-the R of formula (IIb)³It is-H, and-the R of formula (IIb)^3aIt is methyl.

Even further preferably, part-L¹- there is formula (IIb ') structure:

Wherein

Wherein dotted line instruction is by forming amido link connection most on the D of CNP parts nitrogen；

The dotted line instruction marked with asterisk is connected to-L²-；

-R²、-R^2a、-R³、-R^3aWith-X²- used as defined in formula (II)；And

Wherein-L¹- be optionally further substituted, on condition that the not substituted base of the hydrogen marked in formula (IIb ') with asterisk replaces Change.

Preferably, the part-L of formula (IIb ')¹- be not further substituted.

Preferably ,-the X of formula (IIb ')²- it is-C (R⁸R^8a)-。

Preferably ,-the R of formula (IIb ')⁸With-R^8aIndependently selected from-H, methyl and ethyl.It is highly preferred that formula (IIb ')- R⁸With-R^8aIn it is at least one be-H.Even further preferably ,-the R of formula (IIb ')⁸With-R^8aAll it is-H.

Preferably ,-the R of formula (IIb ')²With-R^2aIndependently selected from-H, methyl and ethyl.More preferably, formula (IIb ')- R²With-R^2aIn it is at least one be-H.Even further preferably ,-the R of formula (IIb ')²With-R^2aAll it is H.

Preferably ,-the R of formula (IIb ')³With-R^3aIndependently selected from-H, methyl, ethyl, propyl group and butyl.Even more preferably The ground ,-R of formula (IIb ')³With-R^3aIn it is at least one be methyl.In a same preferred embodiment, formula (IIb ')- R³With-R^3aAll it is-H.It is another also, it is preferred that embodiment in ,-the R of formula (IIb ')³With-R^3aAll it is methyl.

Most preferably ,-the R of formula (IIb ')³It is-H, and-the R of formula (IIb ')^3aIt is methyl.

Preferably, part-L¹- there is formula (IIc) structure：

Wherein dotted line instruction is by forming amido link connection most on the-D of CNP parts nitrogen；And

Wherein-L¹- by-L²- Z or-L²- Z ' substitutes and wherein-L¹- be optionally further substituted, on condition that formula (IIc) The middle hydrogen marked with asterisk is not by-L²- Z or-L²- Z ' or substituent are replaced.

Preferably ,-the L of formula (IIc)¹- by a part-L²- Z or-L²- Z ' substitutes.

Preferably, the part-L of formula (IIc)¹- be not further substituted.

In another preferred embodiment, part-L¹- there is formula (IIc-a) structure:

Preferably ,-the L of formula (IIc-a)¹- by a part-L²- Z or-L²- Z ' substitutes.

Preferably, the part-L of formula (IIc-a)¹- be not further substituted.

In another preferred embodiment, part-L¹- there is formula (IIc-b) structure:

Preferably ,-the L of formula (IIc-b)¹- by a part-L²- Z or-L²- Z ' substitutes.

Preferably, part (moiety)-L of formula (IIc-b)¹- be not further substituted.

Even further preferably, part-L¹- it is selected from formula (IIc-i), (IIc-ii), (IIc-iii), (IIc-iv) and (IIc-v):

Wherein

Unlabelled dotted line instruction is by forming amido link connection most on the-D of CNP parts nitrogen；And

The dotted line instruction marked with asterisk is connected to-L²- Z or-L²-Z’；And

-L¹- be optionally further substituted, on condition that formula (IIc-i), (IIc-ii), (IIc-iii), (IIc-iv) and (IIc-v) the not substituted base of the hydrogen marked in asterisk is replaced.

Preferably, the part-L of formula (IIc-i), (IIc-ii), (IIc-iii), (IIc-iv) and (IIc-v)¹- do not entered One step substitutes.

In a particularly preferred embodiment, part-L¹- be

Wherein

The dotted line instruction marked with asterisk is connected to-L²- Z or-L²-Z’。

Preferably ,-the L of formula (IIc-ii)¹- by a part-L²- Z or-L²- Z ' substitutes.

In a same preferred embodiment, part-L¹- selected from formula (IIc-i '), (IIc-ii '), (IIc-iii '), (IIc-iv ') and (IIc-v '):

Wherein

-L¹- be optionally further substituted, on condition that formula (IIc-i '), (IIc-ii '), (IIc-iii '), (IIc-iv ') The not substituted base of the hydrogen marked in (IIc-v ') with asterisk is replaced.

Preferably, the part-L of formula (IIc-i '), (IIc-ii '), (IIc-iii '), (IIc-iv ') and (IIc-v ')¹- It is not further substituted.

In another particularly preferred embodiment, part-L¹- be

Wherein

Preferably ,-the L of formula (IIc-ii ')¹- by a part-L²- Z or-L²- Z ' substitutes.

In a same preferred embodiment, part-L¹- selected from formula (IIc-i "), (IIc-ii "), (IIc-iii ") (IIc-iv "):

Wherein

-L¹- be optionally further substituted, on condition that formula (IIc-i "), (IIc-ii "), (IIc-iii ") and (IIc-iv ") The not substituted base of the middle hydrogen marked with asterisk is replaced.

Preferably, the part-L of formula (IIc-i "), (IIc-ii "), (IIc-iii ") and (IIc-iv ")¹- not further Substitution.

In another particularly preferred embodiment, part-L¹- be

Wherein

Preferably ,-the L of formula (IIc-ii ")¹- by a part-L²- Z or-L²- Z ' substitutes.

Formula (II), (IIa), (IIb), (IIb '), (IIc), (IIc-i), (IIc-ii), (IIc-iii), (IIc-iv), (IIc-v)、(IIc-i')、(IIc-ii’)、(IIc-iii’)、(IIc-iv’)、(IIc-v’)、(IIc-i”)、(IIc-ii”)、 (IIc-iii) and (IIc-iv ")-L¹- optional be further substituted with base preferably as described above.

Another particularly preferred part-L¹- be recorded in undocumented european patent application 14180004, it corresponds to Shen Please number international application for being PCT/EP2015/067929.Therefore, in another preferred embodiment, part-L¹- have Formula (III) structure：

Wherein

Dotted line instruction is connected to D primary or secondary amine or hydroxyl by forming acid amides or ester connection respectively；

-R¹、-R^1a、-R²、-R^2a、-R³With-R^3aIt is independently selected from-H ,-C (R⁸R^8aR^8b) ,-C (=O) R⁸、-C≡N、- C (=NR⁸)R^8a、-CR⁸(=CR^8aR^8b)、-C≡CR⁸With-T；

-R⁴、-R⁵With-R^5aIt is independently selected from-H ,-C (R⁹R^9aR^9b) and-T；

A1 and a2 is 0 or 1 independently of each other；

Respectively-R⁶、-R^6a、-R⁷、-R^7a、-R⁸、-R^8a、-R^8b、-R⁹、-R^9a、-R^9bBe independently selected from-H, halogen ,-CN ,- COOR¹⁰、-OR¹⁰、-C(O)R¹⁰、-C(O)N(R¹⁰R^10a)、-S(O)₂N(R¹⁰R^10a)、-S(O)N(R¹⁰R^10a)、-S(O)₂R¹⁰、-S (O)R¹⁰、-N(R¹⁰)S(O)₂N(R^10aR^10b)、-SR¹⁰、-N(R¹⁰R^10a)、-NO₂、-OC(O)R¹⁰、-N(R¹⁰)C(O)R^10a、-N (R¹⁰)S(O)₂R^10a、-N(R¹⁰)S(O)R^10a、-N(R¹⁰)C(O)OR^10a、-N(R¹⁰)C(O)N(R^10aR^10b)、-OC(O)N (R¹⁰R^10a)、-T、C_1-20Alkyl, C_2-20Alkenyl and C_2-20Alkynyl；Wherein-T, C_1-20Alkyl, C_2-20Alkenyl and C_2-20Alkynyl optionally quilt One or more identical or different-R¹¹Substitution and wherein C_1-20Alkyl, C_2-20Alkenyl and C_2-20Alkynyl is optionally by one or more It is individual to be interrupted selected from following group：-T-、-C(O)O-、-O-、-C(O)-、-C(O)N(R¹²)-,-S(O)₂N(R¹²)-、-S(O)N (R¹²)-、-S(O)₂-、-S(O)-、-N(R¹²)S(O)₂N(R^12a)-、-S-、-N(R¹²)-、-OC(OR¹²)(R^12a)-、-N(R¹²)C (O)N(R^12a)-and-OC (O) N (R¹²)-；

Respectively-R¹⁰、-R^10a、-R^10bIndependently selected from：-H、-T、C_1-20Alkyl, C_2-20Alkenyl and C_2-20Alkynyl；Wherein-T, C_1-20Alkyl, C_2-20Alkenyl and C_2-20Alkynyl is optionally by one or more identical or different-R¹¹Substitution and wherein C_1-20Alkane Base, C_2-20Alkenyl and C_2-20Alkynyl is optionally interrupted by one or more selected from following group：-T-、-C(O)O-、-O-、-C (O)-、-C(O)N(R¹²)-、-S(O)₂N(R¹²)-、-S(O)N(R¹²)-、-S(O)₂-、-S(O)-、-N(R¹²)S(O)₂N(R^12a)-、- S-、-N(R¹²)-、-OC(OR¹²)(R^12a)-、-N(R¹²)C(O)N(R^12a)-and-OC (O) N (R¹²)-；

Each T-phase is mutually independently selected from phenyl, naphthyl, indenyl, indanyl, tetrahydro naphthyl, C_3-10Cycloalkyl, 3- to 10- Circle heterocycles base and 8- are to 11- 9-membered heterobicyclic bases；Wherein each T is independently optionally by one or more identical or different-R¹¹Substitution；

Respectively-R¹¹It is independently selected from halogen ,-CN, oxo (=O) ,-COOR¹³、-OR¹³、-C(O)R¹³、-C(O)N (R¹³R^13a)、-S(O)₂N(R¹³R^13a)、-S(O)N(R¹³R^13a)、-S(O)₂R¹³、-S(O)R¹³、-N(R¹³)S(O)₂N(R^13aR^13b)、- SR¹³、-N(R¹³R^13a)、-NO₂、-OC(O)R¹³、-N(R¹³)C(O)R^13a、-N(R¹³)S(O)₂R^13a、-N(R¹³)S(O)R^13a、-N (R¹³)C(O)OR^13a、-N(R¹³)C(O)N(R^13aR^13b)、-OC(O)N(R¹³R^13a) and C_1-6Alkyl；Wherein C_1-6Alkyl is optionally by one Individual or multiple identical or different halogen substitutions；

Respectively-R¹²、-R^12a、-R¹³、-R^13a、-R^13bIndependently selected from：- H and C_1-6Alkyl；Wherein C_1-6Alkyl is optionally by one Or multiple identical or different halogen substitutions；

Optionally, one or more-R¹/-R^1a、-R²/-R^2a、-R³/-R^3a、-R⁶/-R^6a、-R⁷/-R^7aPair connect with them The atom connect forms C together_3-10Cycloalkyl or 3- are to 10- circle heterocycles bases；

Optionally, one or more-R¹/-R²、-R¹/-R³、-R¹/-R⁴、-R¹/-R⁵、-R¹/-R⁶、-R¹/-R⁷、-R²/-R³、- R²/-R⁴、-R²/-R⁵、-R²/-R⁶、-R²/-R⁷、-R³/-R⁴、-R³/-R⁵、-R³/-R⁶、-R³/-R⁷、-R⁴/-R⁵、-R⁴/-R⁶、-R⁴/- R⁷、-R⁵/-R⁶、-R⁵/-R⁷、-R⁶/-R⁷Pair ring A is formed together with the atom that they are connected；

A is selected from phenyl；Naphthyl；Indenyl；Indanyl；Tetrahydro naphthyl；C_3-10Cycloalkyl；3- is to 10- circle heterocycles bases；And 8- To 11- 9-membered heterobicyclic bases；

Wherein-L¹- by-L²- Z or-L²- Z ' substitutes and wherein-L¹- be optionally further substituted；

Wherein

-L²- it is single chemical bond or interval body；

- Z is water-solubility carrier；And

- Z ' is water insoluble carrier.

- the L of formula (III)¹- optional be further substituted with base preferably as described above.

Preferably ,-the L of formula (III)¹- by a part-L²- Z or-L²- Z ' substitutes.

In one embodiment ,-the L of formula (III)¹- be not further substituted.

-L¹- other preferred embodiments be disclosed in EP1536334B1, WO2009/009712A1, WO2008/ In 034122A1, WO2009/143412A2, WO2011/082368A2 and US8618124B2, entire contents are by quoting simultaneously Enter herein.

-L¹- other preferred embodiments be disclosed in US8946405B2 and US8754190B2, entire contents pass through It is incorporated herein by reference.Accordingly, it is preferred that part-L¹- there is formula (IV) structure：

Wherein

The dotted line instruction connection most-D of CNP parts and wherein connection are by selected from-OH ,-SH and-NH₂- D What functional group was carried out；

M is 0 or 1；

-R¹With-R²In at least one or both be independently selected from-CN ,-NO₂, optionally substitute aryl, optionally take The heteroaryl in generation, the alkenyl optionally substituted, the alkynyl optionally substituted ,-C (O) R³、-S(O)R³、-S(O)₂R³With-SR⁴,

R¹With-R²In one and only one be selected from-H, the alkyl optionally substituted, the aryl alkyl that optionally substitutes and optionally Substituted heteroaryl alkyl；

-R³The alkyl that selected from-H, optionally substitutes, the aryl optionally substituted, the aryl alkyl that optionally substitutes, optionally substitute Heteroaryl, the heteroaryl alkyl ,-OR optionally substituted⁹With-N (R⁹)₂；

-R⁴Selected from the alkyl optionally substituted, the aryl optionally substituted, the aryl alkyl optionally substituted, optionally substitute it is miscellaneous Aryl and the heteroaryl alkyl optionally substituted；

Respectively-R⁵Independently selected from：- H, the alkyl optionally substituted, the alkenylalkyl optionally substituted, the alkynyl alkane optionally substituted Base, the aryl optionally substituted, the aryl alkyl optionally substituted, the heteroaryl optionally substituted and the heteroaryl alkyl optionally substituted；

-R⁹Selected from-H and the optional alkyl that substitutes；

- Y- is not present, and-X- is-O- or-S-；Or

- Y- is-N (Q) CH₂-, and-X- is-O-；

Q is selected from the alkyl optionally substituted, the aryl optionally substituted, the aryl alkyl optionally substituted, the heteroaryl optionally substituted Base and the heteroaryl alkyl optionally substituted；

Optionally ,-R¹With-R²3 can be joined together to form to 8- yuan of rings；And

Optionally, two-R⁹Heterocycle is formed together with the nitrogen connected with them；

Wherein

-L²- it is single chemical bond or interval body；

- Z is water-solubility carrier；And

- Z ' is water insoluble carrier.

Only in formula (IV) context, the term used has following meanings：

Term " alkyl " used herein include 1 to 8 carbon atom or in some embodiments include 1 to 6 or 1 to Straight chain, side chain or the cyclic saturated hydrocarbon group of 4 carbon atoms.

Term " alkoxy " includes the alkyl closed with oxygen key, including methoxyl group, ethyoxyl, isopropoxy, ring propoxyl group, ring Butoxy etc..

Term " alkenyl " includes the non-aromatic unsaturated hydrocarbons with carbon-to-carbon double bond.

Term " alkynyl " includes the non-aromatic unsaturated hydrocarbons with the key of carbon-to-carbon three.

Term " aryl " includes 6 to 18 carbon, the aromatic hydrocarbyl of preferably 6 to 10 carbon, including such as phenyl, naphthyl and anthracene The group of base (anthracenyl).Term " heteroaryl " includes containing at least one N, O or S atom, comprising 3 to 15 carbon Aromatic ring, at least one N, O or S atom, the aromatic ring for including 3 to 7 carbon are preferably comprised, including such as pyrrole radicals, pyridine radicals, pyrimidine Base, imidazole radicals,It is oxazolyl, differentThe groups such as oxazolyl, thiazolyl, isothiazolyl, quinolyl, indyl, indenyl.

In some cases, alkenyl, alkynyl, aryl or heteroaryl moieties can be by remaining of alkylidene connection and molecule Part connects.In these cases, substituent will be referred to as alkenylalkyl, alkynylalkyl, aryl alkyl or heteroaryl alkyl, table Bright alkylene moiety is connected in alkenyl, alkynyl, aryl or heteroaryl moieties and with the alkenyl, alkynyl, aryl or heteroaryl Between molecule.

Term " halogen " includes bromine, fluorine, chlorine and iodine.

Term " heterocycle " refers to include 3 to 7 carbon atoms and 4 to 8 yuan of aromatics or non-aromatic of at least one N, O or S atom Race's ring.Example is piperidyl, piperazinyl, THP trtrahydropyranyl, pyrrolidines and tetrahydrofuran base and is above-mentioned term " heteroaryl " The Exemplary groups of offer.

When ring system is optionally substituted, suitable substituent is selected from alkyl, alkenyl, alkynyl or other ring, each optionally Ground is further substituted.Optional substituent on any group (including above-mentioned) include halogen, nitro, cyano group ,-OR ,-SR ,- NR₂、-OCOR、-NRCOR、-COOR、-CONR₂、-SOR、-SO₂R、-SONR₂、-SO₂NR₂, wherein each R independently be alkyl, Alkenyl, alkynyl, aryl or heteroaryl, or two R groups form ring together with the atom that they are connected.

Preferably ,-the L of formula (IV)¹- by a part-L²- Z or-L²- Z ' substitutes.

-L¹- other preferred embodiments be disclosed in WO2013/036857A1, be fully incorporated conduct in text With reference to.Accordingly, it is preferred that part-L¹- there is formula (V) structure:

Wherein

The dotted line instruction connection most-D of CNP parts and wherein connection are carried out by-D amine functional group；

-R¹Selected from the C optionally substituted₁-C₆Straight chain, side chain or cyclic alkyl；The aryl optionally substituted；What is optionally substituted is miscellaneous Aryl；Alkoxy；With-NR⁵ ₂；

-R²Selected from-H；The C optionally substituted₁-C₆Alkyl；The aryl optionally substituted；The heteroaryl optionally substituted；

-R³Selected from-H；The C optionally substituted₁-C₆Alkyl；The aryl optionally substituted；The heteroaryl optionally substituted；

-R⁴Selected from-H；The C optionally substituted₁-C₆Alkyl；The aryl optionally substituted；The heteroaryl optionally substituted；

Respectively-R⁵It is independently selected from-H；The C optionally substituted₁-C₆Alkyl；The aryl optionally substituted；Optionally substitute Heteroaryl；Or two-R when connected together⁵Can be cycloalkyl or cycloheteroalkyl；

Wherein

-L²- it is single chemical bond or interval body；

- Z is water-solubility carrier；And

- Z ' is water insoluble carrier.

Only in formula (V) context, the term used has following meanings：

" alkyl ", " alkenyl " and " alkynyl " include 1-8 carbon atom or the straight chain of 1-6 carbon atom or 1-4 carbon atom, Side chain or cyclic hydrocarbon group, wherein alkyl are saturated hydrocarbons, and alkenyl includes one or more carbon-to-carbon double bonds, and alkynyl include one or Multiple keys of carbon-to-carbon three.Unless otherwise indicated, these contain 1-6 C.

" aryl " include 6-18 carbon atom, the aromatic hydrocarbon group of preferably 6-10 carbon atom, including such as phenyl, naphthyl and The group of anthracene." heteroaryl " includes containing at least one N, O or S atom, the aromatic ring for including 3 to 15 carbon, preferably comprises at least One N, O or S atom, the aromatic ring for including 3 to 7 carbon, including as pyrrole radicals, pyridine radicals, pyrimidine radicals, imidazole radicals,Oxazolyl, It is differentThe groups such as oxazolyl, thiazolyl, isothiazolyl, quinolyl, indyl, indenyl.

Term " substituted " refer to comprising one or more substituents replace the alkyl of one or more hydrogen atoms, alkenyl, Alkynyl, aryl or heteroaryl.Substituent can be generally selected from：Halogen, including F, Cl, Br and I；Low alkyl group, including straight chain, branch Chain and ring-type；Low-grade halogenated alkyl, including fluoro-alkyl, chloro alkyl, bromo alkyl and iodo-alkyl；OH；Rudimentary alcoxyl Base, including straight chain, side chain and ring-type；SH；Lower alkylthio, including straight chain, side chain and ring-type；Amino, alkyl amino, two Alkyl amino, silicyl, including aIkylsilyl groups, alkoxysilyl and arylsilyl groups；Nitro；Cyano group；Carbonyl Base；Carboxylic acid, carboxylate, carboxylic acid amide, amino carbonyl；Aminoacyl；Carbamate；Urea；Thiocarbamate；Thiocarbamide； ketne；Sulfone；Sulfonamide；Aryl, including phenyl, naphthyl and anthryl；Heteroaryl, it includes：5- unit's heteroaryls, including pyrroles, miaow Azoles, furans, thiophene,It is azoles, thiazole, differentAzoles, isothiazole, thiadiazoles, triazole,Diazole and tetrazolium, 6- unit's heteroaryls, bag Include pyridine, pyrimidine, pyrazine, and condensed heteroaryl, including benzofuran, benzothiophene, benzoAzoles, benzimidazole, indoles, benzene And thiazole, benzisoxaAzoles and benzisothiazole.

Preferably ,-the L of formula (V)¹- by a part-L²- Z or-L²- Z ' substitutes.

-L¹- another preferred embodiment be disclosed in US7585837B2, entire contents are incorporated herein by reference. Accordingly, it is preferred that part-L¹- there is formula (VI) structure：

Wherein

R¹And R²Independently selected from hydrogen, alkyl, alkoxy, alkoxyalkyl, aryl, alkaryl, aralkyl, halogen, nitre Base ,-SO₃H、-SO₂NHR⁵, amino, ammonium, carboxyl, PO₃H₂And OPO₃H₂；

R³、R⁴And R⁵Independently selected from hydrogen, alkyl and aryl；

Wherein

-L²- it is single chemical bond or interval body；

- Z is water-solubility carrier；And

- Z ' is water insoluble carrier.

The suitable substituent of formula (VI) is alkyl (such as C_1-6Alkyl), alkenyl (such as C_2-6Alkenyl), alkynyl (such as C_2-6Alkynyl), aryl (such as phenyl), miscellaneous alkyl, miscellaneous thiazolinyl, miscellaneous alkynyl, heteroaryl 4 to 7 circle heterocycles of fragrance (such as) or halogen Plain part.

Only in formula (VI) context, the term used has following meanings：

Term " alkyl ", " alkoxy ", " alkoxyalkyl ", " aryl ", " alkaryl " and " aralkyl " represents 1-8 carbon Alkyl such as methyl, ethyl, propyl group, isopropyl and the butyl of atom, preferably 1-4 carbon atom, and the virtue of 6-10 carbon atom Base, such as phenyl and naphthyl.Term " halogen " includes bromine, fluorine, chlorine and iodine.

Preferably ,-the L of formula (VI)¹- by a part-L²- Z or-L²- Z ' substitutes.

-L¹- another preferred embodiment be disclosed in WO2002/089789A1, entire contents are incorporated by reference into Herein.Accordingly, it is preferred that part-L¹- there is formula (VII) structure：

Wherein

L₁It is difunctional linking group,

Y₁And Y₂It is independently O, S or NR⁷；

R²、R³、R⁴、R⁵、R⁶And R⁷Independently selected from hydrogen, C_1-6Alkyl, C_3-12Branched alkyl, C_3-8Cycloalkyl, C_1-6Substitution Alkyl, C_3-8Substituted cycloalkyl, aryl, the aryl of substitution, aralkyl, C_1-6Miscellaneous alkyl, the C of substitution_1-6Miscellaneous alkyl, C_1-6Alcoxyl Base, phenoxy group and C_1-6Miscellaneous alkoxy；

Ar is such part, and when being included in formula (VII), it forms polysubstituted aromatic hydrocarbon or polysubstituted heterocycle Group；

X be chemical bond or by the part in active transport to target cell, hydrophobic part or its combination,

Y is 0 or 1；

Wherein

-L²- it is single chemical bond or interval body；

- Z is water-solubility carrier；And

- Z ' is water insoluble carrier.

Only in formula (VII) context, the term used has following meanings：

Term " alkyl " is understood to include, such as the C of straight chain, side chain substitution_1-12Alkyl, including alkoxy, C_3-8Cycloalkanes Base or substituted cycloalkyl etc..

Term " substituted " is understood to include with one or more not homoatomic additions or replaces functional group or compound In one or more atoms for including.

Substituted alkyl includes carboxyalkyl, aminoalkyl, dialkyl amido, hydroxy alkyl and mercaptoalkyl；Substitution Cycloalkyl includes part (moieties) such as 4- chlorine cyclohexyl；Aryl includes part such as naphthyl；Substituted aryl includes portion Divide the bromo- phenyl of such as 3-；Aralkyl includes part such as toluyl；Miscellaneous alkyl includes part such as ethylthiophene；Substitution Miscellaneous alkyl includes part such as 3- methoxyl groups thiophone；Alkoxy includes part such as methoxyl group；And phenoxy group includes part Such as 3- nitro-phenoxies.Halogen is understood to include including fluorine, chlorine, iodine and bromine.

Preferably ,-the L of formula (VII)¹- by a part-L²- Z or-L²- Z ' substitutes.

In a further preferred embodiment ,-L¹- substructure of formula (VIII) is included,

Wherein

The dotted line instruction marked with asterisk is connected most on the-D nitrogen of CNP parts by forming amido link；

Unlabelled dotted line instruction is connected to-L¹- remainder；And

Wherein

-L²- it is single chemical bond or interval body；

- Z is water-solubility carrier；And

- Z ' is water insoluble carrier.

Preferably ,-the L of formula (VIII)¹- by a part-L²- Z or-L²- Z ' substitutes.

In one embodiment ,-the L of formula (VIII)¹- be not further substituted.

In another preferred embodiment ,-L¹- include the substructure of formula (IX)

Wherein

The dotted line instruction marked with asterisk is connected most on the-D nitrogen of CNP parts by forming amino-formate bond；

Unlabelled dotted line instruction is connected to-L¹- remainder；And

Wherein

-L²- it is single chemical bond or interval body；

- Z is water-solubility carrier；And

- Z ' is water insoluble carrier.

Preferably ,-the L of formula (IX)¹- by a part-L²- Z or-L²- Z ' substitutes.

In one embodiment ,-the L of formula (IX)¹- be not further substituted.

Partly-L¹--D can be connected to by-D any functional group, and preferably, connected by-D amine functional group To-D.This can be N- tenninal amine functional groups or the amine functional group provided by lysine side-chain, i.e., by the lysine of position 4 or 10 There is provided, when CNP has SEQ ID NO:During 1 sequence；There is provided by the lysine of position 7,9,13,14,18 and 24, when CNP has SEQ ID NO:During 38 sequence；There is provided by the lysine of position 8,10,14,15,19 or 25, when CNP has SEQ ID NO:During 25 sequence；There is provided by the lysine of position 9,11,15,16,20 and 26, when CNP has SEQ ID NO:During 24 sequence；With by position 10,12,16,17,21 and 27 lysine offer, when CNP part there is SEQ ID NO:When 23.

In one embodiment, CNP parts are connected to-L by the N- tenninal amine functional groups of CNP parts¹-。

In another embodiment, if CNP parts have SEQ ID NO:1 sequence, CNP parts pass through position 4 Lysine side chain provide amine functional group be connected to-L¹-。

In another embodiment, if CNP parts have SEQ ID NO:1 sequence, CNP parts pass through position 10 Lysine side chain provide amine functional group be connected to-L¹-。

In another embodiment, if CNP parts have SEQ ID NO:25 sequence, CNP parts pass through position 8 Lysine side chain provide amine functional group be connected to-L¹-。

In another embodiment, if CNP parts have SEQ ID NO:25 sequence, CNP parts pass through position The amine functional group that the side chain of 10 lysine provides is connected to-L¹-。

In another embodiment, if CNP parts have SEQ ID NO:25 sequence, CNP parts pass through position The amine functional group that the side chain of 14 lysine provides is connected to-L¹-。

In another embodiment, if CNP parts have SEQ ID NO:25 sequence, CNP parts pass through position The amine functional group that the side chain of 15 lysine provides is connected to-L¹-。

In another embodiment, if CNP parts have SEQ ID NO:25 sequence, CNP parts pass through position The amine functional group that the side chain of 19 lysine provides is connected to-L¹-。

In another embodiment, if CNP parts have SEQ ID NO:25 sequence, CNP parts pass through position The amine functional group that the side chain of 25 lysine provides is connected to-L¹-。

In another embodiment, if CNP parts have SEQ ID NO:24 sequence, CNP parts pass through position 9 Lysine side chain provide amine functional group be connected to-L¹-。

In another embodiment, if CNP parts have SEQ ID NO:24 sequence, CNP parts pass through position The amine functional group that the side chain of 11 lysine provides is connected to-L¹-。

In another embodiment, if CNP parts have SEQ ID NO:24 sequence, CNP parts pass through position The amine functional group that the side chain of 15 lysine provides is connected to-L¹-。

In another embodiment, if CNP parts have SEQ ID NO:24 sequence, CNP parts pass through position The amine functional group that the side chain of 16 lysine provides is connected to-L¹-。

In another embodiment, if CNP parts have SEQ ID NO:24 sequence, CNP parts pass through position The amine functional group that the side chain of 20 lysine provides is connected to-L¹-。

In another embodiment, if CNP parts have SEQ ID NO:24 sequence, CNP parts pass through position The amine functional group that the side chain of 26 lysine provides is connected to-L¹-。

Most preferably, CNP parts have SEQ ID NO:24 sequence, and carried by the side chain of the lysine of position 26 The amine functional group of confession is connected to-L¹-。

It was surprisingly found that compared with the connection in N- ends or the connection of the acyclic portion with CNP (part) ,-L¹- Being connected to significantly reduces affinity of the CNP prodrugs to NPR-B on CNP ring, the reduction reduces the heart again with NPR-B affinity The risk of blood vessel side effect, such as low blood pressure.

Therefore ,-L¹- be preferably conjugated to the-D loop section the side chain of amino acid residue or the loop section of-D Main chain.Even further preferably ,-L¹- covalent the side chain with the amino acid residue for the loop section for being reversibly conjugated to the-D.

The amino acid residue positioned at-D loop section is preferably any amino acid with functional group.

Preferably ,-L¹The amino acid residue of the-- D conjugated with it loop section includes and is selected from following functional group：Carboxylic acid, Primary amine and secondary amine, maleimide, mercaptan, sulfonic acid, carbonic ester, carbamate, hydroxyl, aldehyde, ketone, hydrazine, isocyanates, different sulphur Cyanate, phosphoric acid, phosphonic acids, haloacetyl, alkyl halide, acryloyl group, aryl fluoride, azanol, sulfuric ester, disulphide, ethene Base sulfone, vinyl ketone, diazonium paraffin, oxirane, guanidine and aziridine.Most preferably ,-L¹--the D conjugated with it loop section Amino acid residue include the functional group selected from hydroxyl, primary amine and secondary amine and guanidine.

Partly-L¹--D can be connected to by any kind of connection, on condition that it is reversible.Preferably ,-L¹- logical Cross the connection selected from acid amides, ester, carbamate, acetal, contracting amine aldehyde, imines, oxime, hydrazone, disulphide and acylguanidines be connected to- On D.Even further preferably ,-L¹--D is connected to by the connection selected from acid amides, ester, carbamate and acylguanidines.

In one embodiment ,-L¹- be connected by ester connection with-D.

In another embodiment ,-L¹- be connected by carbamate connection with-D.

In another embodiment ,-L¹- be connected by acylguanidines with-D.

In a preferred embodiment ,-L¹- be connected by acid amides connection with-D.

With-L¹The amino acid residue of-conjugated-D loop section be selected from albumen (proteinogenic) amino acid residue and Non-protein amino acid residue.

In one embodiment, with-L¹The amino acid residue of the-- D being conjugated loop section is non-protein amino acid.

In a preferred embodiment, with-L¹The amino acid residue of the-- D being conjugated loop section is Argine Monohydrochloride. Even further preferably, the amino acid is selected from histidine, lysine, tryptophan, serine, threonine, tyrosine, asparagus fern ammonia Acid, glutamic acid and arginine.Even further preferably, the amino acid is selected from lysine, aspartic acid, arginine and serine. Even further preferably, the amino acid is selected from lysine, arginine and serine.

In one embodiment, with-L¹The amino acid residue of the-- D being conjugated loop section is histidine.It should be understood that institute State histidine and be not present in SEQ ID NO:In 96 sequences and it be only possible to be present in its variant, analog, ortholog thing, In homologue and derivative.

In one embodiment, with-L¹The amino acid residue of the-- D being conjugated loop section is tryptophan.It should be understood that institute State tryptophan and be not present in SEQ ID NO:In 96 sequences and it be only possible to be present in its variant, analog, ortholog thing, In homologue and derivative.

In one embodiment, with-L¹The amino acid residue of the-- D being conjugated loop section is threonine.It should be understood that institute State threonine and be not present in SEQ ID NO:In 96 sequences and it be only possible to be present in its variant, analog, ortholog thing, In homologue and derivative.

In one embodiment, with-L¹The amino acid residue of the-- D being conjugated loop section is tyrosine.It should be understood that institute State tyrosine and be not present in SEQ ID NO:In 96 sequences and it be only possible to be present in its variant, analog, ortholog thing, In homologue and derivative.

In one embodiment, with-L¹The amino acid residue of the-- D being conjugated loop section is glutamic acid.It should be understood that institute State glutamic acid and be not present in SEQ ID NO:In 96 sequences and it be only possible to be present in its variant, analog, ortholog thing, In homologue and derivative.

In one embodiment, with-L¹The amino acid residue of the-- D being conjugated loop section is lysine.Preferably, institute It is in SEQ ID NO to state amino acid:The lysine of 96 position 4, it corresponds in SEQ ID NO:The bad ammonia of 24 position 26 Acid.

In another embodiment, with-L¹The amino acid residue of the-- D being conjugated loop section is aspartic acid.It is preferred that Ground, the amino acid are in SEQ ID NO:The aspartic acid of 96 position 6, it corresponds in SEQ ID NO:24 position 28 Aspartic acid.

In another embodiment, with-L¹The amino acid residue of the-- D being conjugated loop section is arginine.Preferably, The amino acid is in SEQ ID NO:The arginine of 96 position 7, it corresponds in SEQ ID NO:The essence of 24 position 29 Propylhomoserin.

In another embodiment, with-L¹The amino acid residue of the-- D being conjugated loop section is serine.Preferably, The amino acid is in SEQ ID NO:The serine of 96 position 10 or 12.In one embodiment, the amino acid is In SEQ ID NO:The serine of 96 position 10, it corresponds in SEQ ID NO:The serine of 24 position 32.Another In individual embodiment, the amino acid is in SEQ ID NO:The serine of 96 position 12, it corresponds in SEQ ID NO: The serine of 24 position 34.

In a preferred embodiment, with-L¹The amino acid residue of the-- D being conjugated loop section is lysine.Most Preferably ,-D has SEQ ID NO:24 sequence, and-L¹- it is conjugated to the lysine of position 26.

Have also surprisingly found that the length increase of CNP sequences is beneficial for NEP stability：CNP-22 compares CNP-34 NEP degradeds are easier to, and CNP-34 is more susceptible than CNP-38.

In the prodrug of the present invention ,-L²- it is chemical bond or interval body portion.

In one embodiment ,-L²- it is chemical bond.

In another embodiment ,-L²- it is interval body portion.

As-L²- when not being single chemical bond ,-L²- be preferably selected from-T- ,-C (O) O- ,-O- ,-C (O)-,-C (O) N (R^y1)-、-S(O)₂N(R^y1)-、-S(O)N(R^y1)-、-S(O)₂-、-S(O)-、-N(R^y1)S(O)₂N(R^y1a)-、-S-、-N (R^y1)-、-OC(OR^y1)(R^y1a)-、-N(R^y1)C(O)N(R^y1a)-、-OC(O)N(R^y1)-、C_1-50Alkyl, C_2-50Alkenyl and C_2-50Alkynes Base；Wherein-T-, C_1-50Alkyl, C_2-50Alkenyl and C_2-50Alkynyl is optionally by one or more identical or different-R^y2Substitution, and Wherein C_1-50Alkyl, C_2-50Alkenyl and C_2-50Alkynyl is optionally interrupted by one or more selected from following group：-T-、-C(O) O-、-O-、-C(O)-、-C(O)N(R^y3)-、-S(O)₂N(R^y3)-、-S(O)N(R^y3)-、-S(O)₂-、-S(O)-、-N(R^y3)S(O)₂N(R^y3a)-、-S-、-N(R^y3)-、-OC(OR^y3)(R^y3a)-、-N(R^y3)C(O)N(R^y3a)-and-OC (O) N (R^y3)-；

-R^y1With-R^y1aIt is independently selected from-H ,-T, C_1-50Alkyl, C_2-50Alkenyl and C_2-50Alkynyl；Wherein-T, C_1-50Alkane Base, C_2-50Alkenyl and C_2-50Alkynyl is optionally by one or more identical or different-R^y2Substitution, and wherein C_1-50Alkyl, C_2-50 Alkenyl and C_2-50Alkynyl is optionally interrupted by one or more selected from following group：-T-、-C(O)O-、-O-、-C(O)-、-C(O) N(R^y4)-、-S(O)₂N(R^y4)-、-S(O)N(R^y4)-、-S(O)₂-、-S(O)-、-N(R^y4)S(O)₂N(R^y4a)-、-S-、-N (R^y4)-、-OC(OR^y4)(R^y4a)-、-N(R^y4)C(O)N(R^y4a)-and-OC (O) N (R^y4)-；

Each T independently selected from：Phenyl, naphthyl, indenyl, indanyl, tetrahydro naphthyl, C_3-10Cycloalkyl, 3- are miscellaneous to 10- members Ring group, 8- to 11- 9-membered heterobicyclics base, 8- to the more ring groups of 30- member carbon and the miscellaneous more ring groups of 8- to 30- members；Wherein each T is independently optional By one or more identical or different-R^y2Substitution；

Respectively-R^y2Independently selected from：Halogen ,-CN, oxo (=O) ,-COOR^y5、-OR^y5、-C(O)R^y5、-C(O)N (R^y5R^y5a)、-S(O)₂N(R^y5R^y5a)、-S(O)N(R^y5R^y5a),-S(O)₂R^y5、-S(O)R^y5、-N(R^y5)S(O)₂N(R^y5aR^y5b)、- SR^y5、-N(R^y5R^y5a)、-NO₂、-OC(O)R^y5,-N(R^y5)C(O)R^y5a、-N(R^y5)S(O)₂R^y5a、-N(R^y5)S(O)R^y5a、-N (R^y5)C(O)OR^y5a、-N(R^y5)C(O)N(R^y5aR^y5b)、-OC(O)N(R^y5R^y5a) and C_1-6Alkyl；Wherein C_1-6Alkyl is optionally by one Individual or multiple identical or different halogen substitutions；And

Respectively-R^y3、-R^y3a、-R^y4、-R^y4a、-R^y5、-R^y5aWith-R^y5bIndependently selected from：- H and C_1-6Alkyl, wherein C_1-6Alkyl Optionally substituted by one or more identical or different halogens.

As-L²- when not being single chemical bond ,-L²- even more preferably selected from-T- ,-C (O) O- ,-O- ,-C (O)-,-C (O) N (R^y1)-、-S(O)₂N(R^y1)-、-S(O)N(R^y1)-、-S(O)₂-、-S(O)-、-N(R^y1)S(O)₂N(R^y1a)-、-S-、-N (R^y1)-、-OC(OR^y1)(R^y1a)-、-N(R^y1)C(O)N(R^y1a)-、-OC(O)N(R^y1)-、C_1-50Alkyl, C_2-50Alkenyl and C_2-50Alkynes Base；Wherein-T-, C_1-20Alkyl, C_2-20Alkenyl and C_2-20Alkynyl is optionally by one or more identical or different-R^y2Substitution, and Wherein C_1-20Alkyl, C_2-20Alkenyl and C_2-20Alkynyl is optionally interrupted by one or more selected from following group：-T-、-C(O) O-、-O-、-C(O)-、-C(O)N(R^y3)-、-S(O)₂N(R^y3)-、-S(O)N(R^y3)-、-S(O)₂-、-S(O)-、-N(R^y3)S(O)₂N(R^y3a)-、-S-、-N(R^y3)-、-OC(OR^y3)(R^y3a)-、-N(R^y3)C(O)N(R^y3a)-and-OC (O) N (R^y3)-；

-R^y1With-R^y1aIt is independently selected from-H ,-T, C_1-10Alkyl, C_2-10Alkenyl and C_2-10Alkynyl；Wherein-T, C_1-10Alkane Base, C_2-10Alkenyl and C_2-10Alkynyl is optionally by one or more identical or different-R^y2Substitution, and wherein C_1-10Alkyl, C_2-10 Alkenyl and C_2-10Alkynyl is optionally interrupted by one or more selected from following group：-T-、-C(O)O-、-O-、-C(O)-、-C(O) N(R^y4)-、-S(O)₂N(R^y4)-、-S(O)N(R^y4)-、-S(O)₂-、-S(O)-、-N(R^y4)S(O)₂N(R^y4a)-、-S-、-N (R^y4)-、-OC(OR^y4)(R^y4a)-、-N(R^y4)C(O)N(R^y4a)-and-OC (O) N (R^y4)-；

-R^y2Selected from halogen ,-CN, oxo (=O) ,-COOR^y5、-OR^y5、-C(O)R^y5、-C(O)N(R^y5R^y5a)、-S(O)₂N (R^y5R^y5a)、-S(O)N(R^y5R^y5a)、-S(O)₂R^y5、-S(O)R^y5、-N(R^y5)S(O)₂N(R^y5aR^y5b)、-SR^y5、-N (R^y5R^y5a)、-NO₂、-OC(O)R^y5、-N(R^y5)C(O)R^y5a、-N(R^y5)S(O)₂R^y5a、-N(R^y5)S(O)R^y5a、-N(R^y5)C(O) OR^y5a、-N(R^y5)C(O)N(R^y5aR^y5b)、-OC(O)N(R^y5R^y5a) and C_1-6Alkyl；Wherein C_1-6Alkyl is optionally one or more Identical or different halogen substitution；And

Respectively-R^y3、-R^y3a、-R^y4、-R^y4a、-R^y5、-R^y5aWith-R^y5bIt is independently selected from-H and C_1-6Alkyl；Wherein C_1-6 Alkyl is optionally substituted by one or more identical or different halogens.

As-L²- when not being single chemical bond ,-L²- even more preferably selected from-T- ,-C (O) O- ,-O- ,-C (O)-,-C (O) N (R^y1)-、-S(O)₂N(R^y1)-、-S(O)N(R^y1)-、-S(O)₂-、-S(O)-、-N(R^y1)S(O)₂N(R^y1a)-、-S-、-N (R^y1)-、-OC(OR^y1)(R^y1a)-、-N(R^y1)C(O)N(R^y1a)-、-OC(O)N(R^y1)-、C_1-50Alkyl, C_2-50Alkenyl and C_2-50Alkynes Base；Wherein-T-, C_1-50Alkyl, C_2-50Alkenyl and C_2-50Alkynyl is optionally by one or more identical or different-R^y2Substitution, and Wherein C_1-50Alkyl, C_2-50Alkenyl and C_2-50Alkynyl is optionally interrupted by one or more selected from following group：-T-、-C(O) O-、-O-、-C(O)-、-C(O)N(R^y3)-、-S(O)₂N(R^y3)-、-S(O)N(R^y3)-、-S(O)₂-、-S(O)-、-N(R^y3)S(O)₂N(R^y3a)-、-S-、-N(R^y3)-、-OC(OR^y3)(R^y3a)-、-N(R^y3)C(O)N(R^y3a)-and-OC (O) N (R^y3)-；

-R^y1With-R^y1aIndependently selected from-H ,-T, C_1-10Alkyl, C_2-10Alkenyl and C_2-10Alkynyl；

Each T independently selected from：Phenyl, naphthyl, indenyl, indanyl, tetrahydro naphthyl, C_3-10Cycloalkyl, 3- are miscellaneous to 10- members Ring group, 8- to 11- 9-membered heterobicyclics base, 8- to the more ring groups of 30- member carbon and the miscellaneous more ring groups of 8- to 30- members；

Respectively-R^y2Independently selected from：Halogen and C_1-6Alkyl；And

Even further preferably ,-L²- it is C_1-20Alkyl chain, it is optionally independently selected from following group by one or more It is disconnected：- O- ,-T- and-C (O) N (R^y1)-；And the C_1-20Alkyl chain is optionally independently selected from following group by one or more and taken Generation：- OH ,-T and-C (O) N (R^y6R^y6a)；Wherein-R^y1、-R^y6、-R^y6aIndependently selected from H and C_1-4Alkyl, and wherein T is selected from benzene Base, naphthyl, indenyl, indanyl, tetrahydro naphthyl, C_3-10Cycloalkyl, 3- to 10- circle heterocycles base, 8- to 11- 9-membered heterobicyclics base, 8- is to the more ring groups of 30- member carbon and the miscellaneous more ring groups of 8- to 30- members.

Preferably ,-L²- the molecular weight with 14g/mol to 750g/mol.

Preferably ,-L²- include being selected from following part：

Wherein

Dotted line instruction is respectively connecting to-L²-、-L¹- ,-Z and/or-Z' remainder；And

In a preferred embodiment ,-L²- the chain length with 1 to 20 atom.

As used herein, with part-L²- related term " chain length " refers in-L¹- exist between-Z in most short connection - L²- atomicity.

Preferably ,-L²- there is formula (i) structure

Wherein

The dotted line instruction marked with asterisk is connected to-L¹-；

Unlabelled dotted line instruction is connected to-Z or-Z'；

-R¹Selected from-H, C_1-6Alkyl, C_2-6Alkenyl and C_2-6Alkynyl；

N is selected from 0,1,2,3,4,5,6,7,8,9,10,11,12,13,14,15,16,17 and 18；And

The part of wherein formula (i) is optionally further substituted.

Preferably ,-the R of formula (i)¹Selected from-H, methyl, ethyl, propyl group and butyl.Even further preferably ,-the R of formula (i)¹Choosing From-H, methyl, ethyl and propyl group.Even further preferably ,-the R of formula (i)¹Selected from-H and methyl.Most preferably ,-the R of formula (i)¹It is Methyl.

Preferably, the n of formula (i) is selected from 0,1,2,3,4,5,6,7,8,9 and 10.Even further preferably, the n of formula (i) is selected from 0th, 1,2,3,4 and 5.Even further preferably, the n of formula (i) is selected from 0,1,2 and 3.Even further preferably, the n of formula (i) is selected from 0 and 1. Most preferably, the n of formula (i) is 0.

In a preferred embodiment ,-L²- it is to be selected from following part：

Wherein

The dotted line instruction marked with asterisk is connected to-L¹-；

Unlabelled dotted line instruction is connected to-Z or-Z'；And

Which part (ii), (iii), (iv), (v), (vi), (vii), (viii), (ix), (x), (xi), (xii), (xiii), (xiv), (xv), (xvi) and (xvii) is optionally further substituted.

In a preferred embodiment ,-L²- be selected from

Wherein

The dotted line instruction marked with asterisk is connected to-L¹-；And

Unlabelled dotted line instruction is connected to-Z or-Z'.

Even more preferably-L²- be selected from

Wherein

The dotted line instruction marked with asterisk is connected to-L¹-；And

Unlabelled dotted line instruction is connected to-Z or-Z'.

Even further preferably ,-L²- be

Wherein

The dotted line instruction marked with asterisk is connected to-L¹-；And

Unlabelled dotted line instruction is connected to-Z or-Z'.

In a preferred embodiment, part-L¹-L²- be selected from

Wherein

The dotted line instruction marked with asterisk is connected to-Z or-Z '.

In one even more preferably embodiment, part-L¹-L²- be

Wherein

The dotted line instruction marked with asterisk is connected to-Z or-Z '.

In a most preferred embodiment, part-L¹-L²- there is formula (IId-ii ') structure

Wherein

The dotted line instruction marked with asterisk is connected to-Z or-Z '.

In another preferred embodiment, part-L¹-L²- be selected from

Wherein

The dotted line instruction marked with asterisk is connected to-Z or-Z '.

In one even more preferably embodiment, part-L¹-L²- be

Wherein

The dotted line instruction marked with asterisk is connected to-Z or-Z '.

In a most preferred embodiment, part-L¹-L²- there is formula (IId-iia ') structure

Wherein

The dotted line instruction marked with asterisk is connected to-Z or-Z '.

In another preferred embodiment, part-L¹-L²- be selected from

Wherein

The dotted line instruction marked with asterisk is connected to-Z or-Z '.

In one even more preferably embodiment, part-L¹-L²- be

Wherein

The dotted line instruction marked with asterisk is connected to-Z or-Z '.

In a most preferred embodiment, part-L¹-L²- there is formula (IId-iib ') structure

Wherein

The dotted line instruction marked with asterisk is connected to-Z or-Z '.

Partly-L²--L can be connected to by replacing any existing-H¹-。

Preferably, by formula (II)-R¹、-R^1a、-R²、-R^2a、-R³、-R^3a、-R⁴、-R^4a、-R⁵、-R^5a、-R⁶、-R⁷、- R^7a、-R⁸、-R^8a、-R⁹、-R^9a、-R¹⁰、-R^10aAnd/or-R¹¹1 to 5 preferably 1 quilt-L in the hydrogen of offer²- replace.It is preferred that Ground, by formula (III)-R¹、-R^1a、-R²、-R^2a、-R³、-R^3a、-R⁴、-R⁵、-R^5a、-R⁶、-R^6a、-R⁷、-R^7a、-R⁸、-R^8a、- R^8b、-R⁹、-R^9a、-R^9b、-R¹⁰、-R^10a、-R^10b、-R¹¹、-R¹²、-R^12a、-R¹³、-R^13aAnd/or-R^13b1 to 5 in the hydrogen of offer Individual preferably 1 quilt-L²- replace.

Preferably ,-Z has 5 to 200kDa molecular weight.Even further preferably ,-Z is with 8 to 100kDa, it is even more excellent Selection of land 10 is to 80kDa, and even more preferably still 12 to 60, even more preferably still 15 to 40 molecular weight, and most preferably ,-Z tools There is about 20kDa molecular weight.It is another also, it is preferred that embodiment in ,-Z have about 40kDa molecular weight.

Carrier-Z includes C_8-24Alkyl or polymer.Preferably ,-Z includes polymer, is preferably selected from following polymer： 2- methylacryloyls-epoxide ethylphosphocholine, poly- (acrylic acid), poly- (acrylate), poly- (acrylamide), poly- (alcoxyl Base) polymer, poly- (acid amides), poly- (amidoamines), poly- (amino acid), poly- (acid anhydrides), poly- (asparagine), poly- (butyric acid), poly- (glycolic), polybutylene terephthalate, poly- (caprolactone), poly- (carbonic ester), poly- (cyanoacrylate), poly- (dimethyl Acrylamide), poly- (ester), poly- (ethene), PEG, poly- (oxirane), poly- (etherophosphoric acid), poly- (ethylAzoles Quinoline), poly- (glycolic), poly- (dihydroxypropyl ethyl ester), it is poly- (hydroxyethyl-Oxazoline), poly- (hydroxyl-metacrylate), Poly- (hydroxypropylmethacrylamide), poly- (hydroxy propyl methacrylate), poly- (hydroxypropylOxazoline), poly- (imido Base carbonic ester), poly- (lactic acid), polylactic-co-glycolic acid, poly- (Methacrylamide), poly- (methacrylate), poly- (methylOxazoline), poly- (organic phosphine nitrile), poly- (ortho esters), it is poly- (Oxazoline), poly- (propane diols), poly- (siloxanes), poly- (ammonia Carbamate), poly- (vinyl alcohol), poly- (vinyl amine), poly- (vinyl methyl ether), PVP, silicone, Cellulose, carboxymethyl cellulose, HYDROXY PROPYL METHYLCELLULOSE, chitin, chitosan, glucan, dextrin, gelatin, hyaluronic acid With derivative, the hyaluronic acid of functionalization, mannosan, pectin, rhamnose galacturonic acid glycan (rhamnogalacturonan), starch, hydroxyalkyl starch, hydroxy ethyl starch and other polymerizations based on carbohydrate Thing, xylan and its copolymer.

In one embodiment, the water-solubility carrier-Z includes protein.Preferable protein is selected from US 2012/ The carboxy terminal peptide of human chorionic gonadtropin described in 0035101 A1, it is incorporated herein by reference；Albumin；Such as WO XTEN sequences described in 2011123813 A2, it is incorporated herein by reference；As described in the A1 of WO 2011/144756 Proline/alanine random coils sequence, it is incorporated herein by reference；Such as the A1 of WO 2008/155134 and WO 2013/ Proline/alanine/serine random coils sequence described in 024049 A1, it is incorporated herein by reference；Merged with Fc Albumen.

In another preferred embodiment ,-Z includes derivative of fatty acid.Preferable derivative of fatty acid is WO Those disclosed in the 2005/027978 A2 and A1 of WO 2014/060512, it is incorporated herein by reference.

In another preferred embodiment ,-Z is the polymer based on hyaluronic acid.

In one embodiment ,-Z is the carrier disclosed in the A1 of WO 2012/02047, and it is incorporated herein by reference.

In another embodiment ,-Z is the carrier disclosed in the A1 of WO 2013/024048, and it is incorporated by reference into this Text.

In another preferred embodiment ,-Z is the polymer based on PEG.Even further preferably ,-Z be side chain or The polymer based on PEG of multi-arm.Most preferably ,-Z is the polymer based on PEG of multi-arm.Even further preferably ,-Z is more The polymer based on PEG of arm, it has arm of at least four based on PEG.

Preferably, the polymer-Z based on PEG of the side chain or multi-arm, preferably the multi-arm polymer-Z based on PEG It is connected to some-L²-L¹- D, wherein each part-L²-L¹- D is preferably connected to the end of side chain or arm, preferably to arm End.Preferably, the polymer-Z based on PEG of the side chain or multi-arm, preferably the multi-arm polymer-Z connections based on PEG To 2,3,4,5,6,7,8,9,10,11,12,13,14,15 or 16 part-L²-L¹-D.Even further preferably, the side chain or Polymer-the Z based on PEG of multi-arm, preferably the multi-arm polymer-Z based on PEG are connected to 2,3,4,6 or 8 part-L²- L¹-D.Even further preferably, polymer-the Z based on PEG, preferably multi-arm of the side chain or the multi-arm polymerization based on PEG Thing-Z is connected to 2,4 or 6 part-L²-L¹- D, even further preferably, the polymer-Z based on PEG of the side chain or multi-arm, It is preferred that polymer-the Z based on PEG of multi-arm is connected to 4 or 6 part-L²-L¹- D, and most preferably, the side chain or more Polymer-the Z based on PEG of arm, preferably the multi-arm polymer-Z based on PEG are connected to 4 part-L²-L¹-D。

If more than one part-L²-L¹It is favourable that-D, which is connected to a part-Z, as it ensure that sufficiently high medicine Thing loads, and it allows the CNP that pharmacy effective dose is provided with small size, and which again increases the convenience of patient.

Preferable carrier-Z of the water solubility based on PEG is multi-arm PEG derivatives, such as in JenKem Technology, USA product list (can be from http://www.jenkemusa.com/Pages/PEGProducts.aspx, Dec 18,2014 Download and obtain) in be described in detail, such as 4- arm-PEG derivatives, the 4- arm-PEG particularly comprising pentaerythrite core, bag 8- arm-PEG derivatives containing six glycerine cores, and the 8- arm-PEG derivatives containing tripentaerythritol core.It is highly preferred that Carrier-Z of the water solubility based on PEG is included and is selected from following part：

4- arm PEG amine comprising pentaerythrite core：

Wherein n scopes are 20 to 500；

8- arm PEG amine comprising six glycerine cores：

Wherein n scopes are 20 to 500；And

The glycerine of R=six or tripentaerythritol core texture；And

6- arm PEG amine comprising sorbierite or dipentaerythritol core:

Wherein n scopes are 20 to 500；And

R=includes sorbierite or dipentaerythritol core；

And wherein dotted line instruction is connected to the remainder of CNP prodrugs.

The x of formula (Ia) is to be selected from following integer：1st, 2,3,4,5,6,7,8,9,10,11,12,13,14,15 and 16.It is excellent Selection of land, x are to be selected from following integer：2nd, 3,4,6 and 8.It is highly preferred that x is to be selected from following integer：2nd, 4 and 6.It is even more excellent Selection of land, x are to be selected from following integer：4 and 6, and most preferably, x is 4.

The y of formula (Ib) is to be selected from following integer：1st, 2,3,4 or 5.Preferably, y is to be selected from following integer：1st, 2 or 3. In a preferred embodiment, y is 1.In a same preferred embodiment, y is 2.

In another preferred embodiment, a part-L²-L¹- D is connected to a part-Z.

In a particularly preferred embodiment ,-Z is branch polymer.In one embodiment ,-Z is that have one Individual, two, three, four, five or six branch points branch polymer.Preferably ,-Z is with one, two or three The branch polymer of branch point.In one embodiment ,-Z is the branch polymer with a branch point.In another reality Apply in scheme ,-Z is the branch polymer with two branch points.In another embodiment ,-Z is that have three branch points Branch polymer.

Branch point is preferably selected from-N<、-CH<With>C<.

Preferably, the branched fraction-Z is based on PEG.

Preferably, the branched fraction-Z has at least 10kDa molecular weight.

In one embodiment, the branched fraction-Z has 10kDa to 500kDa molecular weight and including end points, More preferably 10kDa is to 250Da and including end points, and even more preferably 10kDa is to 150kDa and including end points, even more preferably 12kDa is to 100kDa and including end points, and most preferably 15kDa is to 80kDa and including end points.

Preferably, the branched fraction-Z has 10kDa to 80kDa molecular weight and including end points.In an implementation In scheme, molecular weight is about 10kDa.In another embodiment, the molecular weight of the branched fraction-Z is about 20kDa. In another embodiment, the molecular weight of the branched fraction-Z is about 30kDa.In another embodiment, the side chain Partly-Z molecular weight is about 40kDa.In another embodiment, the molecular weight of the branched fraction-Z is about 50kDa. In another embodiment, the molecular weight of the branched fraction-Z is about 60kDa.In another embodiment, the side chain Partly-Z molecular weight is about 70kDa.In another embodiment, the molecular weight of the branched fraction-Z is about 80kDa.Most Preferably, the branched fraction-Z has about 40kDa molecular weight.

Applicant have surprisingly discovered that part-L¹-L²- Z N- ends are connected to increase NEP stability aspects than portion internally Position connection is significantly more effective, and on increasing loop section of the most poorly efficient connecting portion in CNP parts of NEP- stability.So And applicant have surprisingly discovered that, by using with least 10kDa, for example, at least 12kDa, for example, at least 15kDa, for example extremely Few 18kDa, for example, at least 20kDa, for example, at least 24kDa, for example, at least 25kDa, for example, at least 27kDa, for example, at least 30kDa Branched fraction-Z can compensate on improve NEP stability ring connection this shortcoming.Preferably, the branched fraction-Z With no more than 500kDa, preferably more than 250kDa, preferably more than 200Da, preferably more than 150kDa and most preferably Molecular weight no more than 100kDa.Most preferably, the branched fraction-Z has about 40kDa molecular weight.Therefore, in CNP portions Point loop section do not only result in increased NEP stability using the branched fraction-Z, and make increased NEP- stability and The NPR-B of the reduction related on ring to being connected to is combined and combined.

Preferably ,-Z or-Z' includes part

In one embodiment ,-Z includes formula (a) part

Wherein

Dotted line instruction is connected to-L²- or to-Z remainder；

BP^aIt is to be selected from following branch point：-N<、-CR<With>C<；

- R is selected from-H and C_1-6Alkyl；

If BP^aIt is-N<Or-CR<, a is 0, and if BP^aIt is>C<, n is 1；

-S^a-、-S^a’-、-S^a”- and-S^a”’- it is chemical bond or selected from C independently of each other_1-50Alkyl, C_2-50Alkenyl and C_2-50Alkynes Base；Wherein C_1-50Alkyl, C_2-50Alkenyl and C_2-50Alkynyl is optionally by one or more identical or different-R¹Substitution, and wherein C_1-50Alkyl, C_2-50Alkenyl and C_2-50Alkynyl is optionally interrupted by one or more selected from following group：-T-、-C(O)O-、- O-、-C(O)-、-C(O)N(R²)-、-S(O)₂N(R²)-、-S(O)N(R²)-、-S(O)₂-、-S(O)-、-N(R²)S(O)₂N (R^2a)-、-S-、-N(R²)-、-OC(OR²)(R^2a)-、-N(R²)C(O)N(R^2a)-and-OC (O) N (R²)-；

Respectively-T- independently selected from：Phenyl, naphthyl, indenyl, indanyl, tetrahydro naphthyl, C_3-10Cycloalkyl, 3- to 10- members Heterocyclic radical, 8- to 11- 9-membered heterobicyclics base, 8- to the more ring groups of 30- member carbon and the miscellaneous more ring groups of 8- to 30- members；Wherein each-T- is independently Optionally by one or more identical or different-R¹Substitution；

Respectively-R¹Independently selected from：Halogen ,-CN, oxo (=O) ,-COOR³、-OR³、-C(O)R³、-C(O)N(R³R^3a)、-S (O)₂N(R³R^3a)、-S(O)N(R³R^3a)、-S(O)₂R³、-S(O)R³、-N(R³)S(O)₂N(R^3aR^3b)、-SR³、-N(R³R^3a)、- NO₂、-OC(O)R³、-N(R³)C(O)R^3a、-N(R³)S(O)₂R^3a、-N(R³)S(O)R^3a、-N(R³)C(O)OR^3a、-N(R³)C(O)N (R^3aR^3b)、-OC(O)N(R³R^3a) and C_1-6Alkyl；Wherein C_1-6Alkyl is optionally taken by one or more identical or different halogens Generation；

Respectively-R²、-R^2a、-R³、-R^3aWith-R^3bIndependently selected from：- H and C_1-6Alkyl, wherein C_1-6Alkyl optionally by one or Multiple identical or different halogen substitutions；And

-P^a’、-P^a”With-P^a”’It is independently polymer moieties.

In one embodiment, the BP of formula (a)^aIt is-N<.

In another embodiment, the BP of formula (a)^aIt is-CR<.Preferably ,-R is-H.Therefore, a of formula (a) is preferably 0。

In another embodiment, the BP of formula (a)^aIt is>C<.

In one embodiment ,-the S of formula (a)^a- it is chemical bond.

In another embodiment ,-the S of formula (a)^a- it is selected from C_1-10Alkyl, C_2-10Alkenyl and C_2-10Alkynyl, the C_1-10Alkane Base, C_2-10Alkenyl and C_2-10Alkynyl is optionally by selected from following one or more chemical groups interruption：-C(O)O-、-O-、-C (O)-、-C(O)N(R⁴)-、-S(O)₂N(R⁴)-、-S(O)N(R⁴)-、-S(O)₂-、-S(O)-、-N(R⁴)S(O)₂N(R^4a)-、- S-、-N(R⁴)-、-OC(OR⁴)(R^4a)-、-N(R⁴)C(O)N(R^4a)-and-OC (O) N (R⁴)-；Wherein-R⁴With-R^4aIndependently select From-H, methyl, ethyl, propyl group and butyl.Preferably ,-the S of formula (a)^a- selected from methyl, ethyl, propyl group, butyl, its optional quilt Interrupted selected from following one or more chemical groups：- O- ,-C (O)-and-C (O) N (R⁴)-。

In one embodiment ,-the S of formula (a)^a’- it is chemical bond.

In another embodiment ,-the S of formula (a)^a’- it is selected from C_1-10Alkyl, C_2-10Alkenyl and C_2-10Alkynyl, the C_1-10Alkane Base, C_2-10Alkenyl and C_2-10Alkynyl is optionally by selected from following one or more chemical groups interruption：-C(O)O-、-O-、-C (O)-、-C(O)N(R⁴)-、-S(O)₂N(R⁴)-、-S(O)N(R⁴)-、-S(O)₂-、-S(O)-、-N(R⁴)S(O)₂N(R^4a)-、- S-、-N(R⁴)-、-OC(OR⁴)(R^4a)-、-N(R⁴)C(O)N(R^4a)-and-OC (O) N (R⁴)-；Wherein-R⁴With-R^4aIndependently select From-H, methyl, ethyl, propyl group and butyl.Preferably ,-the S of formula (a)^a’- selected from methyl, ethyl, propyl group, butyl, its optional quilt Interrupted selected from following one or more chemical groups：- O- ,-C (O)-and-C (O) N (R⁴)-。

In one embodiment ,-the S of formula (a)^a”- it is chemical bond.

In another embodiment ,-the S of formula (a)^a”- it is selected from C_1-10Alkyl, C_2-10Alkenyl and C_2-10Alkynyl, the C_1-10Alkane Base, C_2-10Alkenyl and C_2-10Alkynyl is optionally by selected from following one or more chemical groups interruption：-C(O)O-、-O-、-C (O)-、-C(O)N(R⁴)-、-S(O)₂N(R⁴)-、-S(O)N(R⁴)-、-S(O)₂-、-S(O)-、-N(R⁴)S(O)₂N(R^4a)-、- S-、-N(R⁴)-、-OC(OR⁴)(R^4a)-、-N(R⁴)C(O)N(R^4a)-and-OC (O) N (R⁴)-；Wherein-R⁴With-R^4aIndependently select From-H, methyl, ethyl, propyl group and butyl.Preferably ,-the S of formula (a)^a"-selected from methyl, ethyl, propyl group, butyl, its optional quilt Interrupted selected from following one or more chemical groups：- O- ,-C (O)-and-C (O) N (R⁴)-。

In one embodiment ,-the S of formula (a)^a”’- it is chemical bond.

In another embodiment ,-the S of formula (a)^a”’- it is selected from C_1-10Alkyl, C_2-10Alkenyl and C_2-10Alkynyl, the C_1-10 Alkyl, C_2-10Alkenyl and C_2-10Alkynyl is optionally by selected from following one or more chemical groups interruption：-C(O)O-、-O-、-C (O)-、-C(O)N(R⁴)-、-S(O)₂N(R⁴)-、-S(O)N(R⁴)-、-S(O)₂-、-S(O)-、-N(R⁴)S(O)₂N(R^4a)-、- S-、-N(R⁴)-、-OC(OR⁴)(R^4a)-、-N(R⁴)C(O)N(R^4a)-and-OC (O) N (R⁴)-；Wherein-R⁴With-R^4aIndependently select From-H, methyl, ethyl, propyl group and butyl.Preferably ,-the S of formula (a)^a”’- selected from methyl, ethyl, propyl group, butyl, its optional quilt Interrupted selected from following one or more chemical groups：- O- ,-C (O)-and-C (O) N (R⁴)-。

Preferably ,-the P of formula (a)^a’、-P^a”With-P^a”’Independently include and be selected from following polymer：2- methacryls Base-epoxide ethylphosphocholine, poly- (acrylic acid), poly- (acrylate), poly- (acrylamide), poly- (alkoxy) polymer, poly- (acid amides), poly- (amidoamines), poly- (amino acid), poly- (acid anhydrides), poly- (asparagine), poly- (butyric acid), poly- (glycolic), poly- pair Terephtha-late, poly- (caprolactone), poly- (carbonic ester), poly- (cyanoacrylate), poly- (DMAA), poly- (ester), poly- (ethene), PEG, poly- (oxirane), poly- (etherophosphoric acid), poly- (ethylOxazoline), poly- (glycolic), Poly- (dihydroxypropyl ethyl ester), it is poly- (hydroxyethyl-Oxazoline), poly- (hydroxyl-metacrylate), poly- (hydroxypropylmethyl Acrylamide), poly- (hydroxy propyl methacrylate), poly- (hydroxypropylOxazoline), poly- (iminocarbonic ester), it is poly- (breast Acid), polylactic-co-glycolic acid, poly- (Methacrylamide), poly- (methacrylate), poly- (methylOxazoline), it is poly- (organic phosphine nitrile), poly- (ortho esters), it is poly- (Oxazoline), poly- (propane diols), poly- (siloxanes), poly- (carbamate), poly- (ethene Base alcohol), poly- (vinyl amine), poly- (vinyl methyl ether), PVP, silicone, cellulose, carboxymethyl cellulose Element, HYDROXY PROPYL METHYLCELLULOSE, chitin, chitosan, glucan, dextrin, gelatin, hyaluronic acid and derivative, functionalization Hyaluronic acid, mannosan, pectin, rhamnose galacturonic acid glycan, starch, hydroxyalkyl starch, hydroxy ethyl starch and Other polymer based on carbohydrate, xylan and its copolymers.

It is highly preferred that-the P of formula (a)^a’、-P^a”With-P^a”’Independently include the part based on PEG.Even further preferably, formula (a)-P^a’、-P^a”With-P^a”’The part based on PEG is independently included, it includes at least 20%PEG, even more desirably at least 30%, even more desirably at least 40%PEG, even more desirably at least 50%PEG, even more desirably at least 60%PEG, even more Preferably at least 70%PEG, most preferably at least even more desirably at least 80%PEG and 90%PEG.

Preferably ,-the P of formula (a)^a’、-P^a”With-P^a”’The independently molecular weight with 5kDa to 50kDa and including end points, More preferably there is 5kDa to 40kDa molecular weight and including end points, even more preferably 7.5kDa to 35kDa molecular weight and bag Include end points, even more preferably 7.5 to 30kDa molecular weight, even more preferably 10 to 30kDa molecular weight and including end points.

In one embodiment ,-the P of formula (a)^a’、-P^a”With-P^a”’Molecular weight with about 5kDa.

In another embodiment ,-the P of formula (a)^a’、-P^a”With-P^a”’Molecular weight with about 7.5kDa.

In another embodiment ,-the P of formula (a)^a’、-P^a”With-P^a”’Molecular weight with about 10kDa.

In another embodiment ,-the P of formula (a)^a’、-P^a”With-P^a”’Molecular weight with about 12.5kDa.

In another embodiment ,-the P of formula (a)^a’、-P^a”With-P^a”’Molecular weight with about 15kDa.

In another embodiment ,-the P of formula (a)^a’、-P^a”With-P^a”’Molecular weight with about 20kDa.

In one embodiment ,-Z includes the part of a formula (a).

In another embodiment ,-Z includes the part of two formulas (a).

In another embodiment ,-Z includes the part of three formulas (a).

In another embodiment ,-Z includes the part of four formulas (a).

In another embodiment ,-Z includes the part of five formulas (a).

In another embodiment ,-Z includes the part of six formulas (a).

In a preferred embodiment ,-Z includes the part of two formulas (a).

In a preferred embodiment ,-Z includes the part of formula (b)

Wherein

Dotted line instruction is connected to-L²- or to-Z remainder；

B1 is selected from 0,1,2,3,4,5,6,7 and 8；

B2 is selected from 1,2,3,4,5,6,7 and 8；

B3 is 150 to 1000 integer and including end points；It is preferred that 150 to 500 integer and including end points；And most preferably 200 to 460 integer and including end points；And

B4 is 150 to 1000 integer and including end points；It is preferred that 150 to 500 integer and including end points；And most preferably 200 to 460 integer and including end points.

Preferably, the b3 and b4 of formula (b) are identical integers.

In a preferred embodiment, b3 and b4 is the integer in scope 200 to 250, and most preferably, formula (b) b3 and b4 is about 225.

In another preferred embodiment, b3 and b4 is the integer in scope 400 to 500, and most preferably, The b3 and b4 of formula (b) are about 450.

Preferably, the b1 of formula (b) is selected from 0,1,2,3 and 4.It is highly preferred that the b1 of formula (b) is selected from 1,2 and 3.Most preferably, The b1 of formula (b) is 2.

Preferably, the b2 of formula (b) is selected from 1,2,3,4 and 5.It is highly preferred that the b2 of formula (b) is selected from 2,3 and 4.Most preferably, The b2 of formula (b) is 3.

In an especially preferred embodiment, the b1 of formula (b) is 2, and the b2 of formula (b) is 3, and b3 and b4 are about 450。

In another particularly preferred embodiment, the b1 of formula (b) is 2, and the b2 of formula (b) is 3, and b3 and b4 are about 225。

In one embodiment ,-Z includes the part of a formula (b).

In another embodiment ,-Z includes the part of two formulas (b).

In another embodiment ,-Z includes the part of three formulas (b).

In another embodiment ,-Z includes the part of four formulas (b).

In another embodiment ,-Z includes the part of five formulas (b).

In another embodiment ,-Z includes the part of six formulas (b).

In a preferred embodiment ,-Z includes the part of two formulas (b).

In one even more preferably embodiment ,-Z includes the part of formula (c)

Wherein

Dotted line instruction is connected to-L²- or to-Z remainder；

C1 and c2 is independently 150 to 500 integer and including end points；It is preferred that 200 to 460 integer and including end points.

Preferably, the c1 and c2 of formula (c) are identical integers.

In a preferred embodiment, the c1 and c2 of formula (c) are 200 to 250 and including end points, and most preferably Ground, it is about 225.In another preferred embodiment, the c1 and c2 of formula (c) are 400 to 500 and including end points, and most Preferably, it is about 450.

In a preferred embodiment, part-Z is the branch polymer based on PEG for including at least 10%PEG, With a branch point and two polymeric arms based on PEG, and the molecular weight with about 40kDa.Therefore, two are based on PEG Polymeric arms each have about 20kDa molecular weight.Preferably, branch point is-CH<.

In one embodiment ,-Z includes the part of a formula (c).

In another embodiment ,-Z includes the part of two formulas (c).

In another embodiment ,-Z includes the part of three formulas (c).

In another embodiment ,-Z includes the part of four formulas (c).

In another embodiment ,-Z includes the part of five formulas (c).

In another embodiment ,-Z includes the part of six formulas (c).

In a preferred embodiment ,-Z includes the part of two formulas (c).

In a preferred embodiment, part-Z has formula (d) structure

Wherein

Dotted line instruction is connected to-L²-；

-Z^b- it is selected from C_1-50Alkyl, C_2-50Alkenyl and C_2-50Alkynyl；Wherein C_1-50Alkyl, C_2-50Alkenyl and C_2-50Alkynyl is optional By one or more identical or different-R¹Substitution, and wherein C_1-50Alkyl, C_2-50Alkenyl and C_2-50Alkynyl is optionally by one It is or multiple selected from following group interruption：-T-、-C(O)O-、-O-、-C(O)-、-C(O)N(R²)-、-S(O)₂N(R²)-、-S(O) N(R²)-、-S(O)₂-、-S(O)-、-N(R²)S(O)₂N(R^2a)-、-S-、-N(R²)-、-OC(OR²)(R^2a)-、-N(R²)C(O)N (R^2a)-and-OC (O) N (R²)-；

Respectively-R²、-R^2a、-R³、-R^3aWith-R^3bIndependently selected from：- H and C_1-6Alkyl, wherein C_1-6Alkyl optionally by one or Multiple identical or different halogen substitutions；

And

-Z^aIt is

Wherein

BP^a、-S^a-、-S^a’-、-S^a”-、-S^a”’-、-P^a’、-P^a”、-P^a”’Used with a as being defined to formula (a).

The BP of formula (d)^a、-S^a-、-S^a’-、-S^a”-、-S^a”’-、-P^a’、-P^a”、-P^a”’Preferred embodiment as above face Formula (a) is defined.

In one even more preferably embodiment, part-Z has the structure of formula (e)

Wherein

Dotted line instruction is connected to-L²-；

E is selected from 1,2,3,4,5,6,7,8,9,10,11,12,13,14 and 15；And

-Z^aIt is

Wherein

B1 is selected from 0,1,2,3,4,5,6,7 and 8；

B2 is selected from 1,2,3,4,5,6,7 and 8；

B1, b2, b3 and b4 of formula (e) preferred embodiment are as above defined in face of formula (b).

In one embodiment, the e of formula (e) is 1.In another embodiment, the e of formula (e) is 2.At another In embodiment, the e of formula (e) is 3.In another embodiment, the e of formula (e) is 4.In another embodiment, formula (e) e is 5.In another embodiment, the e of formula (e) is 6.In another embodiment, the e of formula (e) is 7.Another In one embodiment, the e of formula (e) is 8.In another embodiment, the e of formula (e) is 9.In another embodiment, The e of formula (e) is 10.In another embodiment, the e of formula (e) is 11.In another embodiment, the e of formula (e) is 12. In another embodiment, the e of formula (e) is 13.In another embodiment, the e of formula (e) is 14.In another implementation In scheme, the e of formula (e) is 15.

Preferably, the e of formula (e) is selected from 2,3,4,5,6,7,8 and 9.Even further preferably, the e of formula (e) is selected from 3,4,5 and 6.Most preferably, the e of formula (e) is 5.

Preferably, the e of formula (e) is 5, and the b1 of formula (e) is 2, and the b2 of formula (e) is 3, and the b3 and b4 of formula (e) are about 450。

In another preferred embodiment, part-Z is the branch chain polymerization based on PEG for including at least 10%PEG Thing, it has three branch points and 4 polymeric arms based on PEG, and the molecular weight with about 40kDa.Therefore, this four Each of polymeric arms based on PEG has about 10kDa molecular weight.Preferably, each of three branch points is-CH<.

In a preferred embodiment, part-Z has formula (f) structure

Wherein

Dotted line instruction is connected to-L²-；

BP^fIt is to be selected from following branch point：-N<、-CR<With>C<；

- R is selected from-H and C_1-6Alkyl；

If BP^fIt is-N<Or-CR<, f is 0, and if BP^fIt is>C<, f is 1；

-S^f-、-S^f’-、-S^f”- and-S^f”’- it is independently chemical bond or independently selected from C_1-50Alkyl, C_2-50Alkenyl and C_2-50 Alkynyl；Wherein C_1-50Alkyl, C_2-50Alkenyl and C_2-50Alkynyl is optionally by one or more identical or different-R¹Substitute and its Middle C_1-50Alkyl, C_2-50Alkenyl and C_2-50Alkynyl is optionally interrupted by one or more selected from following group：-T-、-C(O)O-、- O-、-C(O)-、-C(O)N(R²)-、-S(O)₂N(R²)-、-S(O)N(R²)-、-S(O)₂-、-S(O)-、-N(R²)S(O)₂N (R^2a)-、-S-、-N(R²)-、-OC(OR²)(R^2a)-、-N(R²)C(O)N(R^2a)-and-OC (O) N (R²)-；

Each R¹Independently selected from：Halogen ,-CN, oxo (=O) ,-COOR³、-OR³、-C(O)R³、-C(O)N(R³R^3a)、-S (O)₂N(R³R^3a)、-S(O)N(R³R^3a)、-S(O)₂R³、-S(O)R³、-N(R³)S(O)₂N(R^3aR^3b)、-SR³、-N(R³R^3a)、- NO₂、-OC(O)R³、-N(R³)C(O)R^3a、-N(R³)S(O)₂R^3a、-N(R³)S(O)R^3a、-N(R³)C(O)OR^3a、-N(R³)C(O)N (R^3aR^3b)、-OC(O)N(R³R^3a) and C_1-6Alkyl；Wherein C_1-6Alkyl is optionally taken by one or more identical or different halogens Generation；

And

-Z^a’、-Z^a‘’With-Z^a”’It is independently

Wherein

The BP of formula (f)^a、-S^a-、-S^a’-、-S^a”-、-S^a”’-、-P^a’、-P^a”With-P^a”’Preferred embodiment as above Formula (a) is defined.

Preferably, the BP of formula (f)²It is-CR<, and r is 0.Preferably ,-R is-H.

Preferably ,-the S of formula (f)^f- it is chemical bond.

Preferably ,-the Z of formula (f)^a’、-Z^a‘’With-Z^a”’With identical structure.Preferably ,-the Z of formula (f)^a’、--Z^a‘’ With-Z^a”’It is formula (b).

Preferably ,-the S of formula (f)^f- it is chemical bond, the BP of formula (f)^aIt is-CR<, wherein-R is-H.Even further preferably, formula (f)-S^f- it is chemical bond, the BP of formula (f)^aIt is-CR<, wherein-R is-H, and-the Z of formula (f)^a’、-Z^a‘’With-Z^a”’It is formula (b) 's.

Even further preferably ,-Z is formula (g)

Wherein

Dotted line instruction is connected to-L²-；

-S^g-、-S^g’- and-S^g”- independently selected from C_1-50Alkyl, C_2-50Alkenyl and C_2-50Alkynyl；Wherein C_1-50Alkyl, C_2-50 Alkenyl and C_2-50Alkynyl is optionally by one or more identical or different-R¹Substitution and wherein C_1-50Alkyl, C_2-50Alkenyl and C_2-50Alkynyl is optionally interrupted by one or more selected from following group：-T-、-C(O)O-、-O-、-C(O)-、-C(O)N (R²)-、-S(O)₂N(R²)-、-S(O)N(R²)-、-S(O)₂-、-S(O)-、-N(R²)S(O)₂N(R^2a)-、-S-、-N(R²)-、-OC (OR²)(R^2a)-、-N(R²)C(O)N(R^2a)-and-OC (O) N (R²)-；

And

-Z^aWith-Z^a‘It is independently

Wherein

The BP of formula (g)^a、-S^a-、-S^a’-、-S^a”-、-S^a”’-、-P^a’、-P^a”With-P^a”’Preferred embodiment as above Formula (a) is defined.

Preferably ,-the S of formula (g)^g- it is selected from C_1-6Alkyl, C_2-6Alkenyl and C_2-6Alkynyl, it is optionally by one or more identical Or difference-R¹Substitution

Wherein

-R¹Selected from halogen, oxo (=O) ,-COOR³、-OR³、-C(O)R³、-C(O)N(R³R^3a)、-S(O)₂N(R³R^3a)、-S (O)N(R³R^3a)、-S(O)₂R³、-S(O)R³、-N(R³)S(O)₂N(R^3aR^3b)、-SR³、-N(R³R^3a)、-NO₂、-OC(O)R³、-N (R³)C(O)R^3a、-N(R³)S(O)₂R^3a、-N(R³)S(O)R^3a、-N(R³)C(O)OR^3a、-N(R³)C(O)N(R^3aR^3b)、-OC(O)N (R³R^3a) and C_1-6Alkyl；Wherein C_1-6Alkyl is optionally substituted by one or more identical or different halogens；And

-R³、-R^3aWith-R^3bIndependently selected from-H, methyl, ethyl, propyl group and butyl.

Even further preferably ,-the S of formula (g)^g- it is selected from C_1-6Alkyl.

Preferably ,-the S of formula (g)^g’- it is selected from C_1-6Alkyl, C_2-6Alkenyl and C_2-6Alkynyl, it is optionally by one or more identical Or difference-R¹Substitution,

Wherein

Even further preferably ,-the S of formula (g)^g’- it is selected from C_1-6Alkyl.

Preferably ,-the S of formula (g)^g”- it is selected from C_1-6Alkyl, C_2-6Alkenyl and C_2-6Alkynyl, it is optionally by one or more identical Or difference-R¹Substitution,

Wherein

Even further preferably ,-the S of formula (g)^g”- it is selected from C_1-6Alkyl.

Preferably ,-the Z of formula (g)^aWith-Z^a‘With identical structure.Preferably ,-the Z of formula (g)^aWith-Z^a‘It is formula (b).

Even further preferably ,-Z is formula (h)

Wherein

Dotted line instruction is connected to-L²-；And

Respectively-Z^cIt is part

Wherein

Each c1 is independently the integer in about 200 to 250 scopes.

Preferably, two c1 of formula (h) are identicals.

Preferably, two c1 of formula (h) are about 225.

In one even more preferably embodiment, part-Z is formula (h-i)

Wherein

Dotted line instruction is connected to-L²-；And

Respectively-Z^cIt is part

Each c1 is independently the integer in 200 to 250 scopes.

Preferably, two c1 of formula (h-i) are identicals.

Preferably, two c1 of formula (h-i) are about 225.

Preferably, CNP prodrugs of the invention are formula (Ia).

Preferably, CNP prodrugs of the invention are formula (Ia), wherein x=1.

In a preferred embodiment, CNP prodrugs of the invention are formula (IIe)

Wherein

The dotted line instruction marked with asterisk is connected to part

Wherein

Each c1 is independently the integer in 400 to 500 scopes.

Preferably, the c1 of formula (IIe) is about 450.

In a same preferred embodiment, CNP prodrugs of the invention are formula (IIe-i)

Wherein

The dotted line instruction marked with asterisk is connected to part

Wherein

Each c1 is independently the integer in 400 to 500 scopes.

Preferably, the c1 of formula (IIe-i) is about 450.

It is another also, it is preferred that embodiment in, CNP prodrugs of the invention are formula (IIe-ii)

Wherein

The dotted line instruction marked with asterisk is connected to part

Wherein

Each c1 is independently the integer in 400 to 500 scopes.

Preferably, the c1 of formula (IIe-ii) is about 450.

In one embodiment, the CNP parts of the CNP prodrugs of formula (IIe), (IIe-i) and (IIe-ii) have SEQ ID NO:25 sequence.

In another embodiment, the CNP parts of the CNP prodrugs of formula (IIe), (IIe-i) and (IIe-ii) have SEQ ID NO:30 sequence.

In a preferred embodiment, the CNP parts of the CNP prodrugs of formula (IIe), (IIe-i) and (IIe-ii) have There are SEQ ID NO:24 sequence.

In one embodiment, CNP part by the nitrogen of CNP N- tenninal amine functional groups be connected to formula (IIe), And-the L in the CNP prodrugs of (IIe-ii) (IIe-i)¹-。

In a preferred embodiment, the nitrogen that the amine functional group for the lysine side-chain that CNP parts pass through CNP provides connects - the L being connected in formula (IIe), (IIe-i) and (IIe-ii) CNP prodrugs¹-。

In one embodiment, if CNP parts have SEQ ID NO:24, the lysine side-chain is not in position 22 And 38 cysteine residues between disulphide bridges formed ring a part (part).

Therefore, in one embodiment, if CNP has SEQ ID NO:24 sequence, CNP parts are by position The amine functional group that the side chain of 9 lysine provides be connected in formula (IIe), (IIe-i) and (IIe-ii) CNP prodrugs- L¹-。

In another embodiment, if CNP has SEQ ID NO:24 sequence, CNP parts are by position 11 The amine functional group that provides of side chain of lysine be connected to-L in formula (IIe), (IIe-i) and (IIe-ii) CNP prodrugs¹-。

In another embodiment, if CNP has SEQ ID NO:24 sequence, CNP parts are by position 15 The amine functional group that provides of side chain of lysine be connected to-L in formula (IIe), (IIe-i) and (IIe-ii) CNP prodrugs¹-。

In another embodiment, if CNP has SEQ ID NO:24 sequence, CNP parts are by position 16 The amine functional group that provides of side chain of lysine be connected to-L in formula (IIe), (IIe-i) and (IIe-ii) CNP prodrugs¹-。

In another embodiment, if CNP has SEQ ID NO:24 sequence, CNP parts are by position 20 The amine functional group that provides of side chain of lysine be connected to-L in formula (IIe), (IIe-i) and (IIe-ii) CNP prodrugs¹-。

In a preferred embodiment, if CNP parts have SEQ ID NO:24, for be connected to formula (IIe), (IIe-i) and the lysine side-chain of the remainder of the CNP prodrugs of (IIe-ii) is residual by the cysteine in position 22 and 38 A part for the ring that disulphide bridges between base are formed.

Therefore, in a preferred embodiment, if CNP has SEQ ID NO:24 sequence, CNP parts pass through Formula (IIe), (IIe-i) and (IIe-ii) CNP prodrugs are connected in the amine functional group that the side chain of the lysine of position 26 provides In-L¹-。

It should be appreciated that the position of above-mentioned cysteine and lysine changes according to the length of CNP parts, and this area Technical staff easily will identify corresponding cysteine and lysine in longer or more short version CNP parts, can also Understand, for example, some lysines may be not present in shorter CNP parts.It should also be understood that the knot as such as direct mutagenesis Fruit, formed in part (part) and there may be more in the acyclic formation part (part) of CNP parts (moiety) and/or ring Lysine residue.

In a preferred embodiment, CNP prodrugs of the invention have formula (IIe), and wherein c1 is about 450, CNP portions Dividing has SEQ ID NO:The 24 sequence and amine functional group provided by the side chain of the lysine in position 26 is connected to-L¹-。

In another preferred embodiment, CNP prodrugs of the invention have formula (IIe-i), and wherein c1 is about 450, CNP parts have SEQ ID NO:24 sequence and the amine functional group connection that the side chain offer of the lysine in position 26 is provided To-L¹-。

In another preferred embodiment, CNP prodrugs of the invention have formula (IIe-ii), and wherein c1 is about 450, CNP parts have SEQ ID NO:24 sequence and the amine functional group connection that the side chain offer of the lysine in position 26 is provided To-L¹-。

Therefore, in a preferred embodiment, CNP prodrugs of the invention have formula (IIe ')

Wherein

Unlabelled dotted line instruction is connected to SEQ ID NO by forming amido link:24 CNP parts in position 26 The nitrogen that the side chain of lysine provides；And

The dotted line instruction marked with asterisk is connected to part

Wherein

Each c1 is independently the integer in 400 to 500 scopes.

Preferably, each c1 of formula (IIe') is about 450.

In another preferred embodiment, CNP prodrugs of the invention have formula (IIe-i ')

Wherein

The dotted line instruction marked with asterisk is connected to part

Wherein

Each c1 is independently the integer in 400 to 500 scopes.

Preferably, each c1 of formula (IIe-i') is about 450.

In another preferred embodiment, CNP prodrugs of the invention have formula (IIe-ii ')

Wherein

The dotted line instruction marked with asterisk is connected to part

Wherein

Each c1 is independently the integer in 400 to 500 scopes.

Preferably, each c1 of formula (IIe-ii') is about 450.

In another preferred embodiment, CNP prodrugs of the invention have formula (IIf)

Wherein

The dotted line instruction marked with asterisk is connected to-the Z with following structure

Wherein

Respectively-Z^aIt is

Wherein

Each c1 is independently the integer in 200 to 250 scopes；Preferably each n is about 225.Preferably, formula (IIf) is each C1 is about 225.

In another preferred embodiment, CNP prodrugs of the invention have formula (IIf-i)

Wherein

Respectively-Z^aIt is

Wherein

Each c1 is independently the integer in 200 to 250 scopes；Preferably each n is about 225.Preferably, formula (IIf-i) Each c1 is about 225.

In another preferred embodiment, CNP prodrugs of the invention have formula (IIf-ii)

Wherein

Respectively-Z^aIt is

Wherein

Each c1 is independently the integer in 200 to 250 scopes；Preferably each n is about 225.Preferably, formula (IIf-ii) Each c1 is about 225.

In one embodiment, the CNP parts of the CNP prodrugs of formula (IIf), (IIf-i) and (IIf-ii) have SEQ ID NO:25 sequence.

In a preferred embodiment, the CNP parts of the CNP prodrugs of formula (IIf), (IIf-i) and (IIf-ii) have There are SEQ ID NO:24 sequence.

In one embodiment, CNP part by the nitrogen of CNP N- tenninal amine functional groups be connected to formula (IIf), And-the L in the CNP prodrugs of (IIf-ii) (IIf-i)¹-。

In a preferred embodiment, the nitrogen that the amine functional group for the lysine side-chain that CNP parts pass through CNP provides connects - the L being connected in formula (IIf), (IIf-i) and (IIf-ii) CNP prodrugs¹-。

In one embodiment, if CNP parts have SEQ ID NO:24, the lysine side-chain is not by position A part for the ring that disulphide bridges between 22 and 38 cysteine residues are formed.

Therefore, in one embodiment, if CNP has SEQ ID NO:24 sequence, CNP parts are by position The amine functional group that the side chain of 9 lysine provides be connected in formula (IIf), (IIf-i) and (IIf-ii) CNP prodrugs- L¹-。

In another embodiment, if CNP has SEQ ID NO:24 sequence, CNP parts are by position 11 The amine functional group that provides of side chain of lysine be connected to-L in formula (IIf), (IIf-i) and (IIf-ii) CNP prodrugs¹-。

In another embodiment, if CNP has SEQ ID NO:24 sequence, CNP parts are by position 15 The amine functional group that provides of side chain of lysine be connected to-L in formula (IIf), (IIf-i) and (IIf-ii) CNP prodrugs¹-。

In another embodiment, if CNP has SEQ ID NO:24 sequence, CNP parts are by position 16 The amine functional group that provides of side chain of lysine be connected to-L in formula (IIf), (IIf-i) and (IIf-ii) CNP prodrugs¹-。

In another embodiment, if CNP has SEQ ID NO:24 sequence, CNP parts are by position 20 The amine functional group that provides of side chain of lysine be connected to-L in formula (IIf), (IIf-i) and (IIf-ii) CNP prodrugs¹-。

In a preferred embodiment, if CNP parts have SEQ ID NO:24, for be connected to formula (IIf), (IIf-i) and the lysine side-chain of the remainder of the CNP prodrugs of (IIf-ii) is residual by the cysteine in position 22 and 38 A part for the ring that disulphide bridges between base are formed.

Therefore, in a preferred embodiment, if CNP has SEQ ID NO:24 sequence, CNP parts pass through Formula (IIf), (IIf-i) and (IIf-ii) CNP prodrugs are connected in the amine functional group that the side chain of the lysine of position 26 provides In-L¹-。

It should be appreciated that the position of above-mentioned cysteine and lysine changes according to the length of CNP parts, and this area Technical staff easily will identify corresponding cysteine and lysine in longer or more short version CNP parts, can also Understand, for example, some lysines may be not present in shorter CNP parts.It should also be understood that the knot as such as direct mutagenesis Fruit, formed in the acyclic formation part of CNP parts and/or ring in part and there may be more lysine residues.

In a preferred embodiment, CNP prodrugs of the invention have formula (IIf), and wherein c1 is about 225, CNP portions Dividing has SEQ ID NO:The 24 sequence and amine functional group provided by the side chain of the lysine in position 26 is connected to-L¹-。

In another preferred embodiment, CNP prodrugs of the invention have formula (IIf-i), and wherein c1 is about 225, CNP parts have SEQ ID NO:24 sequence and the amine functional group connection that the side chain offer of the lysine in position 26 is provided To-L¹-。

In another preferred embodiment, CNP prodrugs of the invention have formula (IIf-ii), and wherein c1 is about 225, CNP parts have SEQ ID NO:24 sequence and the amine functional group connection that the side chain offer of the lysine in position 26 is provided To-L¹-。

In another preferred embodiment, CNP prodrugs of the invention have formula (IIf ')

Wherein

Each Z^aIt is

Wherein

Each c1 is independently the integer in 200 to 250 scopes.

Preferably, each c1 of formula (IIf ') is about 225.

In another preferred embodiment, CNP prodrugs of the invention have formula (IIf-i ')

Wherein

Each Z^aIt is

Wherein

Each c1 is independently the integer in 200 to 250 scopes.

Preferably, each c1 of formula (IIf-i ') is about 225.

In another preferred embodiment, CNP prodrugs of the invention have formula (IIf-ii ')

Wherein

Each Z^aIt is

Wherein

Each c1 is independently the integer in 200 to 250 scopes.

Preferably, each c1 of formula (IIf-ii ') is about 225.

In a word, discovery-L¹- by the way that the side chain of the amino acid in CNP loop section is reversible it is connected to CNP parts, tool There are the branched fraction-Z of at least 10kDa molecular weight application and the combination of the application of the CNP parts more than CNP-22 to cause to have There are the CNP prodrugs of half-life period inside unexpected length.

Carrier-Z ' is insoluble polymer, even more preferably hydrogel.Preferably, the hydrogel include be selected from Under polymer：2- methylacryloyls-epoxide ethylphosphocholine, poly- (acrylic acid), poly- (acrylate), poly- (acryloyl Amine), poly- (alkoxy) polymer, poly- (acid amides), poly- (amidoamines), poly- (amino acid), poly- (acid anhydrides), poly- (asparagine), Poly- (butyric acid), poly- (glycolic), polybutylene terephthalate, poly- (caprolactone), poly- (carbonic ester), poly- (alpha-cyanoacrylate Ester), poly- (DMAA), poly- (ester), poly- (ethene), PEG, poly- (oxirane), poly- (etherophosphoric acid), Poly- (ethylOxazoline), poly- (glycolic), poly- (dihydroxypropyl ethyl ester), it is poly- (hydroxyethyl-Oxazoline), poly- (hydroxyl first Base acrylate), poly- (hydroxypropylmethacrylamide), poly- (hydroxy propyl methacrylate), poly- (hydroxypropylAzoles Quinoline), poly- (iminocarbonic ester), poly- (lactic acid), polylactic-co-glycolic acid, poly- (Methacrylamide), poly- (methyl-prop Olefin(e) acid ester), poly- (methylOxazoline), poly- (organic phosphine nitrile), poly- (ortho esters), it is poly- (Oxazoline), poly- (propane diols), poly- (silica Alkane), poly- (carbamate), poly- (vinyl alcohol), poly- (vinyl amine), poly- (vinyl methyl ether), poly- (ethenyl pyrrolidone Ketone), silicone, cellulose, carboxymethyl cellulose, HYDROXY PROPYL METHYLCELLULOSE, chitin, chitosan, glucan, dextrin, gelatin, Hyaluronic acid and derivative, the hyaluronic acid of functionalization, mannosan, pectin, rhamnose galacturonic acid glycan, starch, hydroxyl Base alkyl starches, hydroxy ethyl starch and other polymer based on carbohydrate, xylan and its copolymers.

If carrier-Z ' is hydrogel, it is preferable that hydrogel includes PEG or hyaluronic acid.Most preferably, the water-setting Glue includes PEG.

Even further preferably, carrier-Z ' is 2011/012715 A1 or WO 2014/ of A2, WO of WO 2006/003014 Hydrogel described in 056926 A1, it is hereby incorporated by reference with full content.

In another embodiment ,-Z' is to assemble the polymer network to be formed by the physics of polymer chain, the physics Aggregation is preferably caused by hydrogen bond, crystallization, spiralization or complexing.In one embodiment, this polymer network is heat setting Xanthan polymer.

Preferably, the gross mass of CNP prodrugs of the invention is at least 10kDa, for example, at least 12kDa, for example, at least 15kDa, for example, at least 20kDa or for example, at least 30kDa.Preferably, the gross mass of CNP prodrugs of the invention is at most 250kDa, such as at most 200kDa, 180kDa, 150kDa or 100kDa.

In a preferred embodiment, the residual activity of CNP prodrugs of the invention is less than 10%, more preferably less than 1%, Even more preferably less than 0.1%, even more preferably less than 0.01%, even more preferably less than 0.001%, more preferably less than 0.0001%.

As used herein, term " residual activity " refers to the activity shown relative to corresponding free CNP, has and carrier knot The activity that the CNP pro-drugs of the invention of the CNP parts of conjunction are shown.Under this linguistic context, term " activity " refers to that NPR-B is tied Close.It should be appreciated that the residual activity of the CNP prodrugs of the measurement present invention needs the time, CNP prodrugs of the present invention will discharge during this period Go out a certain amount of CNP, and measurement results of the CNP of this release by distortion for CNP prodrugs.Therefore, received way It is the conjugate test prodrug being irreversibly stably coupled to wherein drug moiety (being CNP in the case) on carrier Residual activity, it simulates the structure of the CNP prodrugs for measuring residual activity as far as possible.

The A1 of WO 2010/135541, describe in the embodiment 4 of page 143/144 for measuring CNP activity and the present invention The suitable determination method of the residual activity of CNP pro-drugs (preferably stable analogs form).

Another aspect of the present invention is the medicine group comprising at least one CNP prodrugs of the invention and at least one excipient Compound.

In one embodiment, the pharmaceutical composition of the CNP prodrugs comprising the present invention includes the mixing of CNP prodrugs Thing, wherein CNP part are connected to-L by different functional groups¹-, preferably by the amine functional group provided by CNP, that is, pass through N- ends Amine functional group is held, the amine function provided by the side chain of the side chain of the lysine by position 4 and/or the lysine of position 10 Group, when CNP has SEQ ID NO:During 1 sequence；By N- tenninal amine functional groups, by position 8,10,14,15,19 and/ Or the amine functional group that the side chain of 25 lysine provides, when CNP has SEQ ID NO:During 25 sequence；Or pass through N- terminal amines Functional group, the amine functional group provided by the side chain of the lysine in position 9,11,15,16,20 and/or 26, when CNP has SEQ ID NO:During 24 sequence.

In a preferred embodiment, the CNP parts of all CNP prodrugs included in pharmaceutical composition are logical The identical amine functional group for crossing CNP offers is connected to-L¹-, that is, pass through N- tenninal amine functional groups or the lysine in position 4 The amine functional group that the side chain of the amine functional group that side chain provides or the lysine in position 10 provides, as the sequence SEQ that CNP has ID NO:When 1；The amine functional group provided by the side chain of the lysine in position 8,10,14,15,19 or 25, when CNP has SEQ ID NO:During 25 sequence；Or the amine function for the side chain offer for passing through the lysine in position 9,11,15,16,20 or 26 Group, when CNP has SEQ ID NO:During 24 sequence.Most preferably, all CNP prodrugs included in pharmaceutical composition CNP parts-L is connected to by identical amine functional group¹-, the amine functional group is by the amine of the side chain offer of lysine 26 Functional group, when CNP parts have SEQ ID:During NO 24 sequence.

Preferably, comprising the present invention at least one CNP prodrugs pharmaceutical composition pH scopes for pH 3 to pH 8 simultaneously Including end points.It is highly preferred that the pH scopes of pharmaceutical composition be pH 4 to pH6 and including end points.Most preferably, the drug regimen The pH scopes of thing are pH 4 to pH 5 and including end points.

In one embodiment, the medicine group comprising at least one CNP prodrugs of the invention and at least one excipient Compound is liquid or suspension formulations.It should be appreciated that if the CNP prodrugs of the present invention include water insoluble carrier-Z', medicine Composition is suspension formulations.

In another embodiment, the medicine comprising at least one CNP prodrugs of the invention and at least one excipient Composition is drying agent.

The liquid, suspension or dry pharmaceutical compositions include at least one excipient.Used in parenteral administration Excipient can be divided into such as buffer, isotonic modifying agent, preservative, stabilizer, anti-adsorbent, oxidation protection agent, tackifier/viscous Spend reinforcing agent or other adjuvants.However, in some cases, a kind of excipient can have dual or triple functions.Preferably, At least one of pharmaceutical composition included in present invention excipient is selected from：

(i) buffer：The buffer that the physiology of pH holdings within the required range is resistant to, such as sodium phosphate, bicarbonate Salt, succinate, histidine, citrate and acetate, sulfate, nitrate, chloride, acetonate；It can also use Antiacid such as Mg (OH)₂Or ZnCO₃；

(ii) isotonic modifying agent：Reducing can the pain caused by cellular damage caused by permeable pressure head at injection reservoir； Glycerine and sodium chloride are examples；Valid density can be oozed by osmosis using the hypothesis of 285-315mOsmol/kg serum Pressure concentration determines thoroughly；

(iii) preservative and/or antimicrobial：Multiple dose parenteral administration needs to add the preservative of enough concentration To reduce the risk of injection postoperative infection patient to greatest extent, and corresponding management is established and has required；Typical preservative includes Metacresol, phenol, methyl p-hydroxybenzoate, ethyl-para-hydroxybenzoate, propylparaben, P-hydroxybenzoic acid fourth Ester, methaform, benzylalcohol, phenylmercuric nitrate, thimerosal (thimerosol), sorbic acid, potassium sorbate, benzoic acid, chloreresol and benzene Prick oronain.

(iv) stabilizer：By strengthening protein stabilizing power, by making denatured state unstable, or by the way that excipient is straight Connect and be attached to protein to realize stabilisation；Stabilizer can be amino acid, such as alanine, arginine, aspartic acid, sweet ammonia Acid, histidine, lysine, proline, sugar such as glucose, sucrose, trehalose, polyalcohol such as glycerine, mannitol, sorbierite, salt Such as potassium phosphate, sodium sulphate, chelating agent such as EDTA, six phosphate, part bivalent metal ion (zinc, calcium etc.), other salt Or organic molecule such as amphyl；In addition it is possible to use oligomer or polymer for example cyclodextrin, glucan, dendritic macromole, PEG or PVP or nucleoprotamine or HAS；

(v) anti-adsorbent：Mainly use ion or nonionic surfactant or other protein or soluble polymer To coat or compete it is adsorbed onto the inner surface of the container of preparation；Such as poloxamer (Pluronic F-68), PEG dodecanes Base ether (Brij 35), polysorbate20 and 80, glucan, polyethylene glycol, PEG- polyhistidines, BSA and HSA and gelatin； The concentration and type of the excipient of selection depend on the effect to be avoided, but are generally being formed just above CMC value in interface Surfactant mono-layer；

(vi) oxidation protection agent：Antioxidant, such as ascorbic acid, Ectoin (ectoine), methionine, paddy Guang Sweet peptide, MTG, morin, polyethyleneimine (PEI), propylgallate and vitamin E；Chelating agent can also be used Such as citric acid, EDTA, six phosphoric acid and TGA；

(vii) tackifier or viscosity intensifier：Sedimentation of the particle in bottle and syringe is prevented, and for promotion Grain mixing and be suspended again and make suspension be easier to injection (that is, the low power in syringe plunger)；Suitable tackifier are viscous It is such as carbomer tackifier such as Carbopol 940, Carbopol Ultrez 10, cellulose derivative such as hydroxyl to spend reinforcing agent Propyl methocel (hydroxypropyl methylcellulose, HPMC) or DEAE-cellulose (DEAE or DEAE-C), colloidal silicon Sour magnesium (Veegum) or sodium metasilicate, hydroxyapatite gel, tricalcium phosphate gel, xanthans, carrageenan such as Satia GumUTC 30, aliphatic poly (carboxylic acid) for example poly- (D, L- or Pfansteihl) (PLA) and poly- (glycolic) (PGA) and they Copolymer (PLGA), D, L- lactides, the terpolymer of glycolide and caprolactone, poloxamer, hydrophily are poly- (oxygen ethene) Block and poly- (oxypropylene) block of hydrophobicity with form the three block of poly- (oxygen ethene)-poly- (oxypropylene)-poly- (oxygen ethene) (such as), polyetherester copolymer such as polyethylene terephthalate/polybutylene terephthalate copolymer, sugarcane Sugared acetate isobutyrate (SAIB), glucan or derivatives thereof, the combination of glucan and PEG, dimethyl silicone polymer, collagen, Chitosan, polyvinyl alcohol (PVA) and derivative, poly- alkylimide, poly- (acrylamide -co- diallyl dimethyl ammonium (DADMA)), polyvinylpyrrolidone (PVP), glycosaminoglycan (GAGs) such as dermatan sulfate, chondroitin sulfate, keratosulfate Element, heparin, Heparan sulfate, hyaluronic acid, by hydrophobicity A blocks (such as polylactide (PLA) or poly- (lactide-co- Glycolide) (PLGA)) and the ABA three blocks or AB of hydrophilic B block such as polyethylene glycol (PEG) or polyvinylpyrrolidone composition Block copolymer；This block copolymer and above-mentioned poloxamer can show reverse Thermogelling behavior (at room temperature Fluid state is in order to apply, the gel state after injection more than body temperature sol-gel transition temperature)；

(viii) diffusant or dispersant：Connective is modified by hydrolyzing the component of extracellular matrix in space in the cell The permeability of tissue, such as, but not limited to hyaluronic acid, the polysaccharide found in the space between cells of connective tissue；Diffusant, example As but be not limited to hyaluronidase temporarily reduce the viscosity of extracellular matrix and promote inject medicine diffusion；With

(ix) other adjuvants：Such as wetting agent, viscosity modifier, antibiotic, hyaluronidase；Bronsted lowry acids and bases bronsted lowry such as hydrochloric acid and hydrogen Adjuvant necessary to pH regulations during sodium oxide molybdena is manufacture.

Another aspect of the present invention is CNP prodrugs or its pharmaceutically acceptable salt or at least one for including the present invention Purposes of the pharmaceutical composition of CNP prodrugs as medicine.

Another aspect of the present invention is CNP prodrugs or its pharmaceutically acceptable salt or comprising at least one of the invention The pharmaceutical composition of CNP prodrugs, it, which is used to treat, can use the method for the CNP diseases treated.

Preferably, the disease is selected from achondroplasia, osteochondrodysplasia, of short and small stature, nanism, bone cartilage hair Educate bad (osteochondrodysplasias), thanatophoric dysplasia, osteogenesis imperfecta (osteogenesis Imperfecta), achondrogenesis (achondrogenesis), punctatae (chondrodysplasia Punctata), homozygote achondroplasia (homozygous achondroplasia), camptomelic dysplasia (camptomelic dysplasia), congenital lethal hypophosphatasia, perinatal period lethal type osteogenesis imperfecta, short rib are more Refer to syndrome, limb root type punctatae, Jansen (Jansen) type metaphyseal dysplasia (metaphyseal Dysplasia), spondyloepiphyseal dysplasia congenita (spondyloepiphyseal dysplasia congenita), bone It is hypoplasia disease (atelosteogenesis), diastrophic dysplasia (diastrophic dysplasia), congenital short Bone development in femur, Lange (Langer) type mesomelic dysplasia (mesomelic dysplasia), Nievergelt type limbs Bad, Robinow syndromes, Reinhardt syndromes, acrodysostosis (acrodysostosis), peripheral bone development Obstacle (peripheral dysostosis), Kniest depauperations, fibrocartilage occur (fibrochondrogenesis), Roberts syndromes, acra mesomelic dysplasia (acromesomelic dysplasia), manomelia disease (micromelia), Morquio syndromes, Kniest syndromes, metatrophic dysplasia, vertebra epiphysis metaphyseal dysplasia (spondyloepimetaphyseal dysplasia), neurofibromatosis (neurofibromatosis), Legius synthesis Sign, LEOPARD syndromes, Noonan syndromes, hereditary gingival fibromatosis (hereditary gingival Fibromatosis), 1 type neurofibromatosis, Legius syndromes, Cardiofaciocutaneous syndromes, Costello syndromes, SHOX deficiency diseases, idiopathic short stature, growth hormone deficiency, osteoarthritis, skull lock bone development Bad (cleidocranial dysostosis), craniosynostosis (such as Muenke syndromes, Crouzon syndromes, Apert Syndrome, Jackson-Weiss syndromes, Pfeiffer syndromes or Crouzonodermoskeletal syndromes), refer to/toe Type (dactyly), brachydactylia/toe (brachydactyly), camptodactylia/toe (camptodactyly), polydactyly (polydactyly), zygodactyly (syndactyly), Dyssegmental depauperations, enchondrosis, fiber sexual development Bad, hereditary multiple exostoses (hereditary multiple exostoses), hypophosphatemic rickets (hypophosphatemic rickets), Jaffe-Lichtenstein syndromes, Marfan syndromes, McCune- Albright syndromes, osteopetrosis and osteopoikilosis.

Preferably, the disease is selected from achondroplasia, osteochondrodysplasia, of short and small stature, nanism, bone cartilage hair Educate bad, thanatophoric dysplasia, osteogenesis imperfecta, achondrogenesis, punctatae, homozygote cartilage development not Entirely, camptomelic dysplasia, congenital lethal hypophosphatasia, perinatal period lethal type osteogenesis imperfecta, short rib refer to synthesis more Disease, limb root type punctatae, Jansen type metaphyseal dysplasia, spondyloepiphyseal dysplasia congenita, bone development Bone in insufficiency disorder, diastrophic dysplasia, congenital short femur, Lange type mesomelic dysplasia, Nievergelt type limbs Depauperation, Robinow syndromes, Reinhardt syndromes, acrodysostosis, peripheral dysostosis, Kniest hairs Educate bad, fibrocartilage generation, Roberts syndromes, acra mesomelic dysplasia, manomelia disease, Morquio syndromes, Kniest syndromes, metatrophic dysplasia, vertebra epiphysis metaphyseal dysplasia, neurofibromatosis, Legius are comprehensive Simulator sickness, LEOPARD syndromes, Noonan syndromes, hereditary gingival fibromatosis, 1 type neurofibromatosis, Legius synthesis Sign, Cardiofaciocutaneous syndromes, Costello syndromes, SHOX deficiency diseases, idiopathic short stature, growth swash Plain deficiency disease and osteoarthritis.

In another embodiment, the disease is ophthalmology disease, such as glaucoma and/or elevated intraocular pressure.

In another embodiment, the disease is relevant with the overactivity of FGFR3 in cancer, such as multiple marrow Knurl, myeloproliferativesyndromes, leukaemia, plasma cell leukemia, lymthoma, spongioblastoma, prostate cancer, carcinoma of urinary bladder or breast Gland cancer.

In another embodiment, the disease is vascular smooth myopathy, is preferably selected from hypertension, ISR, artery Hardening, acute decompensated heart failure, congestive heart failure, cardiac edema, nephredema, edema due to dysfunction of the liver are swollen, acute renal Incomplete and chronic renal insufficiency.

Preferably, the disease is to be selected from following achondroplasia phenotype：Growth retardation, craniofacial deformities, correction lack Sunken, neck marrow compressing, spinal stenosis, hydrocephalus, the hearing loss as caused by chronic otitis, angiocardiopathy, sacred disease and fertilizer It is fat.

Most preferably, the disease is achondroplasia.

In one embodiment, the patient for undergoing subject treatment method is mammalian subject, preferably human patientses. In one embodiment, the human patientses are adults.In a preferred embodiment, human patientses are that paediatrics is suffered from Person.

Another aspect of the present invention is CNP prodrugs or its pharmaceutically acceptable salt or at least one for including the present invention The pharmaceutical composition of CNP prodrugs prepare be used for treat can with CNP treat disease medicine in purposes.

Preferably, the disease is selected from achondroplasia, osteochondrodysplasia, of short and small stature, nanism, bone cartilage hair Educate bad, thanatophoric dysplasia, osteogenesis imperfecta, achondrogenesis, punctatae, homozygote cartilage development not Entirely, camptomelic dysplasia, congenital lethal hypophosphatasia, perinatal period lethal type osteogenesis imperfecta, short rib refer to synthesis more Disease, limb root type punctatae, Jansen type metaphyseal dysplasia, spondyloepiphyseal dysplasia congenita, bone development Bone in insufficiency disorder, diastrophic dysplasia, congenital short femur, Lange type mesomelic dysplasia, Nievergelt type limbs Depauperation, Robinow syndromes, Reinhardt syndromes, acrodysostosis, peripheral dysostosis, Kniest hairs Educate bad, fibrocartilage generation, Roberts syndromes, acra mesomelic dysplasia, manomelia disease, Morquio syndromes, Kniest syndromes, metatrophic dysplasia, vertebra epiphysis metaphyseal dysplasia, neurofibromatosis, Legius are comprehensive Simulator sickness, LEOPARD syndromes, Noonan syndromes, hereditary gingival fibromatosis, 1 type neurofibromatosis, Legius synthesis Sign, Cardiofaciocutaneous syndromes, Costello syndromes, SHOX deficiency diseases, idiopathic short stature, growth swash Plain deficiency disease, osteoarthritis, cleidocranial dysostosis, craniosynostosis (such as Muenke syndromes, Crouzon syndromes, Apert syndromes, Jackson-Weiss syndromes, Pfeiffer syndromes or Crouzonodermoskeletal syndromes), Finger/toe type, brachydactylia/toe, camptodactylia/toe, polydactyly, zygodactyly, Dyssegmental depauperations, enchondrosis, fibre Tie up sexual organ dysplasia, hereditary multiple exostoses, hypophosphatemic rickets, Jaffe-Lichtenstein synthesis Sign, Marfan syndromes, McCune-Albright syndromes, osteopetrosis and osteopoikilosis.

Most preferably, the disease is achondroplasia.

In one embodiment, with CNP prodrugs or its pharmaceutically acceptable salt or include the present invention at least one The disease of the medicine composite for curing of CNP prodrugs occurs in mammalian subject, preferably in human patientses.In an implementation In scheme, the human patientses are adults.In a preferred embodiment, human patientses are pediatric patients.

Another aspect of the present invention is to need to treat and can suffered from the mammal of the CNP one or more diseases treated The method for being treated in the preferred human patientses of person, controlling, postpone or preventing, including it is effective to the patient therapeuticallv in need The step of CNP prodrugs or its pharmaceutically acceptable salt of amount or pharmaceutical composition comprising CNP prodrugs of the present invention.In a reality Apply in scheme, human patientses are adults.In a preferred embodiment, human patientses are pediatric patients.

Preferably, the one or more diseases that can be treated with CNP are selected from achondroplasia, osteochondrodysplasia, body Material is short and small, nanism, osteochondrodysplasia, thanatophoric dysplasia, osteogenesis imperfecta, achondrogenesis, point-like cartilage hair It is lethal to educate incomplete, homozygote achondroplasia, camptomelic dysplasia, congenital lethal hypophosphatasia, perinatal period Type osteogenesis imperfecta, short rib refer to syndrome, limb root type punctatae, Jansen type metaphyseal dysplasia, congenital more Bone development in centrum epiphysis dysplasia, Atelosteogenesis, diastrophic dysplasia, congenital short femur, Lange type limb Bad, Nievergelt types mesomelic dysplasia, Robinow syndromes, Reinhardt syndromes, acrodysostosis, week Enclosing property dysostosis, Kniest depauperations, fibrocartilage generation, Roberts syndromes, acra mesomelic dysplasia, Manomelia disease, Morquio syndromes, Kniest syndromes, metatrophic dysplasia, vertebra epiphysis metaphyseal dysplasia, Neurofibromatosis, Legius syndromes, LEOPARD syndromes, Noonan syndromes, hereditary gingival fibromatosis, 1 type god Through fibromatosis, Legius syndromes, Cardiofaciocutaneous syndromes, Costello syndromes, SHOX deficiency diseases, (such as Muenke is comprehensive for idiopathic short stature, growth hormone deficiency, osteoarthritis, cleidocranial dysostosis, craniosynostosis Simulator sickness, Crouzon syndromes, Apert syndromes, Jackson-Weiss syndromes, Pfeiffer syndromes or Crouzonodermoskeletal syndromes), refer to/toe type, brachydactylia/toe, camptodactylia/toe, polydactyly, zygodactyly, Dyssegmental depauperations, enchondrosis, fibrous dysplasia, hereditary multiple exostoses, hypophosphate Hypophosphatemic rickets, Jaffe-Lichtenstein syndromes, Marfan syndromes, McCune-Albright syndromes, bone are hard Change disease and osteopoikilosis.

In another embodiment, it with the CNP one or more diseases treated can be ophthalmology disease, such as glaucoma And/or elevated intraocular pressure.

In another embodiment, one or more diseases that CNP is treated and the excessive work of FGFR3 in cancer can be used Change relevant, such as Huppert's disease, myeloproliferativesyndromes, leukaemia, plasma cell leukemia, lymthoma, spongioblast Knurl, prostate cancer, carcinoma of urinary bladder or breast cancer.

In another embodiment, it can be vascular smooth myopathy with the CNP one or more diseases treated, preferably select From hypertension, ISR, artery sclerosis, acute decompensated heart failure, congestive heart failure, cardiac edema, renal plevis product Water, edema due to dysfunction of the liver are swollen, acute renal insufficiency and chronic renal insufficiency.

Preferably, can be to be selected from following achondroplasia phenotype with the CNP one or more diseases treated：Growth Slow, craniofacial deformities, correction defect, the compressing of neck marrow, spinal stenosis, hydrocephalus, the hearing loss as caused by chronic otitis, painstaking effort Pipe disease, sacred disease and obesity.

Most preferably, can be achondroplasia with the CNP one or more diseases treated.

Another aspect of the present invention is the pharmaceutical composition using CNP prodrugs, its pharmaceutically acceptable salt or the present invention Method, wherein methods described include using CNP prodrugs, its pharmaceutically acceptable salt or the present invention pharmaceutical composition step Suddenly, described apply passes through following progress：Local, enteral or parenteral administration or applications, the method for injection or infusion, including Intra-articular, periarticular, intracutaneous, subcutaneous, intramuscular, in intravenous, bone, in intraperitoneal, intrathecal, intracapsular, socket of the eye, in vitreum, In tympanum, in bladder, in heart, under transtracheal, subcutaneous, capsule, under arachnoid, in backbone, intra-ventricle, breastbone inner injection and defeated Note, is directly passed by the implanted device (such as Ommaya Reservoir) for allowing for present invention etc. to be transported to brain tissue or brain liquid Brain, direct intraventricular injection or infusion are delivered to, injects or is infused into brain or brain relevant range, be expelled to space under choroid, socket of the eye Injection and eyes instil afterwards, preferably pass through hypodermic injection.

In a preferred embodiment, the present invention relates to CNP prodrugs or its pharmaceutically acceptable salt or the present invention Pharmaceutical composition, it is used for subcutaneous injection treatment achondroplasia.

Another aspect of the present invention is formula (IVa) and the irreversible conjugate of (IVb)：

Wherein

- D is CNP parts；

-L²- it is single chemical bond or interval body portion；

- Z is water-solubility carrier part；

Y is to be selected from following integer：1st, 2,3,4 and 5.

-D、-L²- ,-Z, x and y preferred embodiment it is as described above.

Fig. 1:According to SEQ ID NO:1 CNP structure.

Embodiment

Material and method

CNP SEQ ID No:1 is obtained from Bachem AG, Bubendorf, Switzerland (CNP-22, the mankind, catalog number (Cat.No.) H- 1296).CNP-34SEQ ID No:40 and CNP-38SEQ ID No:24 derive from CASLO ApS, Kongens Lyngby, red Wheat.

The side chain protected on TCP resins of the Lys26 side chains of N- ends and ivDde protections with Boc protections CNP-38 (being synthesized by Fmoc strategies) derives from CASLO ApS, Kongens Lyngby, Denmark.

The TCP Tentagel of Lys12, Lys16 or Lys22 side chain of N- ends and ivDde protections with Boc protections The CNP-34 (being synthesized by Fmoc strategies) of side chain protected on resin is obtained from Peptide Specialty Laboratories GmbH, Heidelberg, Germany.Side chain protected on TCP tentagel resins with free N-terminal CNP-38 (by The synthesis of Fmoc strategies) Peptide Specialty Laboratories GmbH, Heidelberg are derived from, Germany.

Methoxyl group PEG amine 5kDa derives from Rapp Rapp Polymere GmbH, Tuebingen, Germany.Make in this work Every other PEG obtains from NOF Europe N.V., Grobbendonk, Belgium.

FmocN-Me-Asp (OtBu)-OH derives from Bachem AG, Bubendorf, Switzerland.S- trityl -6- sulfydryls oneself Acid is purchased from Polypeptide, Strasbourg, France.HATU is obtained from Merck Biosciences GmbH, Schwalbach/ Ts, Germany.

2,4- dimethyl benzyl alcohols derive from abcr GmbH, Karlsruhe, Germany.

Fmoc-N-Me-Asp (OBn)-OH derives from Peptide International Inc., Louisville, KY, USA。

Neutral endopeptidase (NEP) derives from Enzo Life Sciences GmbH,Germany.

Every other chemicals and reagent are purchased from Sigma Aldrich GmbH, Taufkirchen, Germany.

Be equipped with polyethylene frit syringe (MultiSynTech GmbH, Witten, Germany) be used as reaction vessel or Washing step for peptide resin.

The general procedure of ivDde protection groups is removed on the CNP of side chain protected from resin

By resin in DMF pre-swollen 30 minutes, discard solvent.By using (v/v, the 2.5mL/g tree of DMF/ hydrazine hydrates 4/1 Fat) it is incubated resin 8x 15min removing ivDde groups.For each step, fresh DMF/ hydrazine hydrate solutions are used.Finally, Resin is washed with DMF (10x), DCM (10x) and is dried in vacuo.

RP-HPLC is purified：

For preparative RP-HPLC, using the controllers of Waters 600 and 2487 dual absorption photometric detectors, it is equipped with following Post：Waters XBridge^TM5 μm, 150x 10mm, flow velocity 6mL/min or Waters XBridge of BEH300Prep C18^TM 10 μm, 150x 30mm, flow velocity 40mL/min of BEH300Prep C18.Using solvent system A (containing 0.1%TFA v/v or 0.01% dense HCl v/v water) and solvent system B (acetonitrile containing the dense HCl v/v of 0.1%TFA v/v or 0.01%) line Property gradient.

If not stated otherwise, merge the HPLC fractions containing product, and be freeze-dried.

Flash chromatography

Using Biotage KP-Sil silicagel columns and normal heptane and ethyl acetate as eluant, eluent, in Sweden Biotage Purification by flash chromatography is carried out in AB Isolera One systems.Product detects in 254nm.

Analysis method

The super performance LC of analytic type is carried out in the Waters Acquity systems equipped with Waters BEH300C18 posts (UPLC)-MS (2.1 × 50mm, 1.7 μm of granularities, flow velocity：0.25mL/min；Solvent orange 2 A：Water containing 0.04%TFA (v/v), it is molten Agent B：Acetonitrile containing 0.05%TFA (v/v)), the Waters Acquity systems and Thermo Scientific LTQ Orbitrap Discovery mass spectrographs are combined or combined with Waters Micromass ZQ.

If not stated otherwise, using equipped with the 5/150GL posts (Amersham of Superdex 200 Bioscience/GE Healthcare) (it is equipped with 0.45 μm of inlet filter) Amersham Bioscience AEKTAbasic systems carry out SEC (SEC).Using 20mM sodium phosphates, 140mM NaCl, pH 7.4 as stream Dynamic phase.

Due to-L¹The reversible nature of-extremely-D connection, it is steady to carry out NEP using the stable analogs of the CNP prodrugs of the present invention The qualitative and measure of receptor affinity, i.e., they are that with similar structure, (it is with stable using the CNP prodrugs to the present invention Connection rather than-Z to-D can reverse connection) carry out.

This is necessary, because the CNP prodrugs of the present invention will discharge CNP, and the CNP of the release in experimentation Result will be influenceed.

The total CNP-38 concentration of blood plasma quantifies

Pass through N- end tags peptide (signature peptide) (sequence after quantitative Trypsin Induced： ) and C- end tag peptide (sequences LQEHPNAR：IGSMSGLGC) the total CNP-38 concentration of blood plasma is determined.

Pass through by using with the united Agilent 1290UPLC of Agilent 6550iFunnel Q-TOF mass spectrographs ESI probes carry out LC-MS analyses.In Waters Acquity BEH300C18 analytical columns (50 × 2.1mm with prefilter I.D., 1.7 μm of granularities) on chromatography carried out with 0.25mL/min (T=25 DEG C) flow velocity.Using containing 0.2% formic acid (v/ V) the acetonitrile of water (UPLC grades) as mobile phase A and containing 0.2% formic acid (UPLC levels) is used as Mobile phase B.Gradient system It is included in the short isocratic step of 3.0 minutes at initial parameter 0.1%B, was then linearly increasing in 17 minutes from 0.1%B 16%B.Mass spectral analysis, monitoring ion m/z482.75 [M+2H] are carried out under single ion monitoring (SIM) pattern²⁺(N- ends) and m/z 824.36[M+H]¹⁺(C- ends).Internal standard is used as using deuterated CNP-38 peptides.

The calibration standard items of CNP-38 conjugates are prepared as follows in blank plasma：First by the Li- heparin machins of defrosting Blood plasma is homogenized, and is then centrifuged for 5 minutes.CNP-38 conjugate formulations are diluted to 10 μ g/mL (the conjugate CNP-38 in DMSO Equivalent) working solution, and (conjugate CNP-38 works as with 9.3ng/100 μ L (conjugate CNP-38 equivalents) and 139.5ng/100 μ L Amount) between concentration add blank plasma in.These solution are used to produce calibration curve.For two kinds of labelled peptides (N- and C- ends End), calibration curve weighting 1/x².For quality control, three Quality control samples are correspondingly prepared for, its content is 116.2ng/100 μ L (high QC, conjugate CNP-38 equivalent), 69.75ng/100 μ L (middle QC, conjugate CNP-38 equivalent) and 23.25ng/100 μ L (low QC, conjugate CNP-38 equivalent).

For sample preparation, protein is carried out by adding 300 μ L precoolings (0 DEG C) methanol into 100 μ L plasma sample Precipitation.200 μ L of supernatant liquid are transferred in new orifice plate and are evaporated to dryness (under 35 DEG C of gentle nitrogen stream).Use 100 μ L Reconstruct solvent (Thermo digestion buffer solutions, job number 60109-101, Thermo Fisher Scientific GmbH, Dreieich, Germany) dissolving residue.By 20 μ g trypsase (job number V5111, Promega GmbH, Mannheim, moral State) it is dissolved in 20 μ L 10mM acetic acid.2 μ L trypsin solutions are added into each cavity.

After being incubated 4 hours under 37 DEG C (water-bath), 5 μ L 0.5M TCEP solution is added to each cavity, and at 96 DEG C It is incubated again 5 minutes.After sample is cooled to room temperature, 3 μ L acetonitriles are added.Eluent is transferred in bottle.10 μ L are expelled to In UPLC-MS systems.

Embodiment 1

Connector reagent 1f synthesis

According to following flow synthesis connector reagent 1f：

To N- methyl-N-Boc- ethylenediamines (2g, 11.48mmol) and NaCNBH₃The MeOH of (819mg, 12.63mmol) 2,4,6- TMBs (2.08g, 10.61mmol) are added in (20mL) solution by several times.Mixture is stirred in rt 90min, it is acidified with 3M HCl (4mL) and is stirred for 15min.Reactant mixture is added into saturation NaHCO₃In solution (200mL), And use CH₂Cl₂Extract 5x.By the organic phase Na of merging₂SO₄Dry, and solvent is evaporated in vacuo.By obtained N- methyl- N-Boc-N '-Tmob- ethylenediamines 1a is dried in high vacuum, is not further purified in next reactions steps.

Yield:3.76g (11.48mmol, 89% purity, 1a:Product=8 of double Tmob protections:1)

MS:M/z 355.22=[M+H]⁺, (single isotopic mass=354.21. of calculating

To 1a (2g, 5.65mmol) CH₂Cl₂In (24mL) solution, COMU (4.84g, 11.3mmol), N-Fmoc- are added N-Me-Asp (OBn)-OH (2.08g, 4.52mmol) and 2,4,6- trimethylpyridines (2.65mL, 20.34mmol).Reaction is mixed Compound stirs 3h in rt, uses CH₂Cl₂(250mL) dilutes, and with 0.1M H₂SO₄(100mL) washs 3x, and uses salt solution (100mL) washs 3x.By aqueous phase CH₂Cl₂(100mL) is extracted again.By the organic phase Na of merging₂SO₄Dry, filtering simultaneously will Residue is concentrated into 24mL volumes.Using purified by flash chromatography 1b.

Yield:5.31g (148%, 6.66mmol)

MS:M/z 796.38=[M+H]⁺, (single isotopic mass=795.37 of calculating)

Into 1b (5.31g, for the most 4.52mmol of N-Fmoc-N-Me-Asp (OBn)-OH) THF (60mL) solution, Add DBU (1.8mL, 3%v/v).Solution is stirred into 12min in rt, uses CH₂Cl₂(400mL) dilutes and with 0.1M H₂SO₄ (150mL) washs 3x, and washs 3x with salt solution (150mL).By aqueous phase CH₂Cl₂(100mL) is extracted again.By the organic phase of merging Use Na₂SO₄Dry and filter.Separation 1c is further purified after evaporation solvent uses in next reaction.

MS:M/z 574.31=[M+H]⁺, (single isotopic mass=573.30 of calculating)

1c (5.31g, 4.52mmol, crude product) is dissolved in acetonitrile (26mL), addition COMU (3.87g, 9.04mmol), 6- trityls mercaptohexanoic acid (2.12g, 5.42mmol) and 2,4,6- trimethylpyridines (2.35mL, 18.08mmol).Will reaction Mixture stirs 4h in rt, uses CH₂Cl₂(400mL) dilutes and with 0.1M H₂SO₄(100mL) washs 3x, and uses salt solution (100mL) washs 3x.By aqueous phase CH₂Cl₂(100mL) is extracted again.By the organic phase Na of merging₂SO₄Dry, filter, evaporation 1d is separated after solvent.Using purified by flash chromatography product 1d.

Yield:2.63g (62%, 94% purity)

MS:M/z 856.41=[M+H]⁺, (single isotopic mass=855.41 of calculating)

To 1d (2.63g, 2.78mmol) i-PrOH (33mL) and H₂In O (11mL) solution add LiOH (267mg, 11.12mmol), and by reactant mixture in rt 70min is stirred.By mixture CH₂Cl₂(200mL) dilutes and uses 0.1M H₂SO₄(50mL) washs 3x, and washs 3x with salt solution (50mL).By aqueous phase CH₂Cl₂(100mL) is extracted again.By having for merging Machine mutually uses Na₂SO₄Dry, filtering, and 1e is separated after evaporation solvent.Using purified by flash chromatography 1e.

Yield:2.1g (88%)

MS:M/z 878.4=[M+Na]⁺, (single isotopic mass=837.40 of calculating)

DCC (123mg, 0.59mmol) is added into 1e (170mg, 0.198mmol) anhydrous DCM (4mL) solution and is urged The DMAP of change amount.After 5min, add N- hydroxy-succinimides (114mg, 0.99mmol) and stir reactant mixture in rt 1h.Reactant mixture is filtered, solvent is removed in a vacuum and takes residue is molten in 90% acetonitrile+0.1%TFA (3.4mL) In.Crude mixture is purified by RP-HPLC.Product fraction is neutralized and concentrated with the phosphate buffers of 0.5M pH 7.4.Will Remaining aqueous phase is extracted with DCM, and 1f is separated after evaporation solvent.

Yield:154mg (81%)

MS:M/z 953.4=[M+H]⁺, (single isotopic mass=952.43 of calculating)

Embodiment 2

N^εK4/εK10- CNP list-connectors mercaptan 2, N^εK4- CNP list-connector mercaptan 2c and N^εK10- CNP list-connector sulphur Alcohol 2d synthesis

N is prepared by the way that CNP-22 (5.2 μm of ol) is dissolved in 0.6mL DMSO^εK4/εK10- CNP list-connectors mercaptan (tool Have the mixture of the region isomer of the connector for the side-chain amino group for being conjugated in Lys4 or Lys10) 2.Add 0.15mL 0.375M Borate buffer solution, adjusted to pH 8.5, added in 60 μ L DIPEA and 0.34mL DMSO with TBAH hydrate 1f (6.1mg, 7.1 μm of ol), mixture is stirred at room temperature 30 minutes.Reactant mixture is with 2mL acetonitrile/waters 1/1 (v/v) Diluted with 200 μ L AcOH, separate the N of protection from reactant mixture by RP-HPLC^εK4/εK10- CNP list-connectors are conjugated Thing.

The RP-HPLC gradients of optimization can be used for separating N^εK4- CNP list-connector mercaptan 2a and N^εK10- CNP list-connectors Mercaptan 2b.

By using the lyophilized product of 0.6mL 90/10/2/2 (v/v/v/v) HFIP/TFA/TES/ water process at room temperature Fraction 1 hour realizes the removal of blocking group.De-protected N is purified by RP-HPLC^εK4/εK10- CNP list-connector mercaptan 2.The identity and purity of product are determined by ESI-LCMS.

The N of deprotection^εK4- CNP list-connector mercaptan 2c and N^εK10- CNP list-connector mercaptan 2d can respectively from 2a and 2b is similarly obtained.

Embodiment 3

N^αG1The synthesis of-CNP list-connectors mercaptan 3

By dissolving CNP-22 (5.2 μm of ol) N is prepared in 0.6mL DMSO^αG1- CNP list-connectors mercaptan 3.Add The 0.25mL 0.5M phosphate buffers pH 7.4 and 1f (6.1mg, 7.1 μm of ol) in 0.34mL DMSO, and by mixture A few hours are stirred in rt.Reactant mixture is diluted with 2mL acetonitrile/waters 1/1 (v/v) and 200 μ L AcOH, and passes through RP-HPLC The N of protection is separated from reactant mixture^αG1- CNP list-connector mercaptan.

By using the lyophilized product of 0.6mL 90/10/2/2 (v/v/v/v) HFIP/TFA/TES/ water process at room temperature Fraction 1 hour realizes the removal of blocking group.De-protected N is purified by RP-HPLC^αG1- CNP list-connectors mercaptan 3.It is logical Cross identity and purity that ESI-LCMS determines product.

Embodiment 4

CNP is mono--connector mercaptan 2c, 2d and 3 PEGylation

By 1 μm of ol CNP it is mono--connector mercaptan 2c is dissolved in 0.5mL acetonitriles/0.2M Succinate Buffers pH 3.8 1/ In 1 (v/v), the linear 40kDa PEG- maleimides of 1.2 μm of ol are added, and mixture is stirred in rt.By adding 20 μ Reaction is quenched L AcOH, and purifies CNP conjugates 4 by preparing RP-HPLC.

CNP conjugates 5 and 6 by 1 μm of ol CNP it is mono--connector mercaptan 2d and 3 is prepared as.

CNP contents are determined by the quantitative amino acid analysis after all hydrolyzing in acid condition.

Embodiment 5

Vitro release kinetics

CNP conjugates 4,5 and 6 are dissolved in 60mM sodium phosphates, 3mM EDTA with about 2mg/mL concentration, 0.01% told In temperature -20, pH 7.4, and it is sterile filtered.Mixture is incubated at 37 DEG C.Aliquot is taken out at time point and passes through RP- HPLC and ESI-MS analyses.Map the UV- signal integration related to the CNP of release and relative to incubation time.

Application curves fitting software estimates to discharge half-court (halftime of release) accordingly.

Embodiment 6

Pharmacokinetics and cGMP generations in rat

Iv and isodose CNP-22, CNP conjugate 4,5 or 6 is injected with sc in normal rat.The blood elapsed over time Starch the horizontal determinations (B2 of United States Patent (USP) 8,377,884) as described in the literature of CNP and cGMP.

Embodiment 7

The synthesis of the 6- mercaptohexanoic acids 7 of Dmb protections

Compound 7 is synthesized according to following flow:

Into 6- mercaptohexanoic acids (7.10g, 47.90mmol) trifluoroacetic acid (20mL) solution, 2,4- dimethylbenzyls are added Alcohol (13.5g, 95.80mmol).Mixture is stirred into 60min in RT, then removes trifluoroacetic acid in a vacuum.Residue is molten Solution stirs 60min in 95.8mL LiOH (3M) and THF (81mL) mixture, and in rt.Solvent is removed in a vacuum and is incited somebody to action Water-based residue extracts 3x with EtOAc (200mL).By the organic phase MgSO of merging₄Dry, and remove solvent in a vacuum. 7 are purified by RP-HPLC.

Yield:2.27g (8.52mmol, 18%)

MS:M/z 267.01=[M+H]⁺, (single isotopic mass=266.13 of calculating)

Embodiment 8

Connector reagent 8c synthesis

According to following flow synthesis connector reagent 8c:

Into 1c (21.6g, 27.18mmol) isopropanol (401mL) solution add water (130mL) and LiOH (3.90g, 163.06mmol).Reactant mixture is stirred into 3h in rt, then by it with toluene (300mL) dilution, and with 0.1M HCl (200mL) washs 3x.The aqueous phase of merging is washed into 3x with toluene (100mL).Aqueous phase 4M NaOH (4mL) are alkalized to pH 8.5 and use CH₂Cl₂(200mL) extracts 8x.By the CH of merging₂Cl₂Mutually washed with salt solution (50mL), use Na₂SO₄Dry.Evaporate molten 8b is separated after agent, is not further purified and is used in next reaction.

Yield:11.89g (24.59mmol, 90%)

MS:M/z 484.16=[M+H]⁺, (single isotopic mass=483.26 of calculating)

In N₂Under-atmosphere, to 7 (293mg, 1.10mmol) and PyBOP (572mg, 1.10mmol) THF (10mL) solution Middle addition DIEA (0.52mL, 3.00mmol).Reactant mixture is stirred into 60min in rt.Add 8b (484mg, 1.00mmol) THF (2mL) solution, and reaction is stirred for 60min.Reaction is quenched with 2M citric acid solutions (10mL) and removed under vacuo Remove THF.Then obtained aqueous phase is extracted into 2x with EtOAc (15mL), and by the organic layer of merging water (10mL) and salt solution (10mL) is washed, and uses MgSO₄Dry.Solvent is removed in a vacuum, and 8c is purified by RP HPLC.

Yield:330mg (0.451mmol, 45%)

MS:M/z 732.34=[M+H]⁺, (single isotopic mass=731.38 of calculating)

Embodiment 9

The synthesis of connector reagent 9

According to following flow synthesis connector reagent 9:

Add into 8b (2.00g, 4.14mmol) and Fmoc-Cl (1.07g, 4.14mmol) dioxane (20mL) solution Enter 1M Na₂CO₃Solution (20mL).Reactant mixture is stirred into 40min in rt.Water (100mL) and ether (100mL) are added, and Aqueous phase is extracted into 2x with ether (100mL).Aqueous phase is acidified until pH 1 with dense HCl, and extracts 3x with ether again.It will merge Organic phase Na₂SO₄Dry, and remove solvent in a vacuum.It is not further purified and uses 9 in the next step.

Yield:2.63g (3.73mmol, 90%)

MS:M/z 728.32=[M+Na]⁺, (single isotopic mass=705.33 of calculating)

Embodiment 10

Reversible Lys26CNP-38PEG2x20kDa conjugates 10f synthesis

According to following flow synthesis conjugate 10f:

On the TCP resins that 2.00g (0.21mmol) is had to the N- ends of Boc protections and the Lys26 side chains of ivDde protections The CNP-38 of side chain protected remove ivDde protections according to the program that is provided in material and method, obtain 10a.Connector is tried Agent 8c (336mg, 0.46mmol), PyBOP (239mg, 0.46mmol) and DIEA (182 μ L, 1.04mmol) DMF (5mL) are molten Liquid is incubated 10min under rt, is then added in resin 10a.Suspension is shaken at room temperature 2 hours.By residue DMF (10mL) washs 10x, washs 10x with DCM (10mL), is dried in vacuo 15min.It is mixed that (- 18 DEG C) cracking are precooled by using 15mL Compound 68.5/10/10/5/3.5/1 (v/w/v/v/v/v) TFA/DTT/ THIOANISOLEs/phenol/water/TIPS processing resins are real Show peptide from the cracking of resin and the removal of blocking group.Mixture is warming up to room temperature and stirs 60min.By thick 10c in precooling Precipitated in ether (- 18 DEG C).Sediment is dissolved in ACN/ water, and is purified by RP-HPLC.The HPLC fractions of merging is direct For next step.

MS:M/z 1124.60=[M+4H]⁴⁺,([M+4H]⁴⁺Calculating single isotopic mass=1124.59)

In the HPLC fractions (250mL) of merging to 10c, add 40mL 0.5M citrate buffer solutions (pH=5.00) and 1/1 (v/v) acetonitrile/water solution of the 7mL sulphur of 0.01M 2,2 '-two double (pyridine-N-oxides).5min is incubated at room temperature Afterwards, reaction is completed.Use the 500mL water containing 0.1%TFA (v/v) to dilute in mixture, and about pH is acidified to AcOH (20mL) 2.10d is purified by RP-HPLC.

Yield:101mg (17.3 μm of ol, 9%) CNP-38- connectors-Dmb*10TFA

MS:M/z 1124.10=[M+4H]⁴⁺,([M+4H]⁴⁺Calculating single isotopic mass=1124.09).

By cleavage mixture 100/5/3/2/1 (v/v/w/v/v) TFA/MSA/DTT/ for adding 30mL precoolings (- 18 DEG C) Water/THIOANISOLE stirs 3 hours to 10d (101mg, 17.3 μm of ol) at 0 DEG C, to realize the cracking of Dmb blocking groups. Thick 10e is precipitated in precooling (- 18 DEG C) ether.Sediment is dissolved in the water containing 0.1%TFA (v/v), and is incubated 10min is to hydrolyze any TFA esters.10e is purified by RP-HPLC.

Yield:46mg (8.34 μm of ol, 48%) CNP-38- connectors-mercaptan * 10TFA

MS:M/z 1094.58=[M+4H]⁴⁺,([M+4H]⁴⁺Calculating single isotopic mass=1094.57)

PEG is added into 10e (46mg, the 8.43 μm of ol) solution in the 1.15mL water containing 0.1%TFA (v/v) 2x20kDa maleimides (GL2-400MA, 870mg, 21.75 μm of ol of Sunbright) contain 0.1%TFA in 4.35mL (v/v) solution in water, 0.5M lactic acid buffers (1.07mL, pH=4.20) are then added.Mixture is stirred at room temperature 4 Hour.Pass through RP-HPLC purified conjugation things 10f.

Yield:233mg (5.21 μm of ol, 62%) conjugate 10f*10HCl

Embodiment 11

Reversible Lys26CNP-38PEG4x10kDa conjugate conjugates 11i synthesis

According to following flow synthesis conjugate 11i:

DIEA is added into 9 (353mg, 0.50mmol) and PyBOP (260mg, 0.50mmol) DMF (9mL) solution (105μL,0.60mmol).By the mixture add to Lys26- side chains deprotection CNP-38 resins 10a (2.00g, 0.21mmol), and by suspension shake at room temperature 2 hours, obtain resin 11a.Resin is washed 10 times with DMF (7mL).With DMF (47mL) solution of HOBt (0.68g, 5.03mmol) and piperazine (3.00g, 34.83mmol) carries out Fmoc protection groups in 11a Cracking.Therefore, resin is incubated together with 10mL cleavage mixture 5 times, carries out 15min in rt every time.Then, DMF is used (7mL) washs resin 7 times.

Prepare Fmoc-Lys (Fmoc)-OH (449mg, 0.76mmol), COMU (325mg, 0.76mmol) and DIEA (165 μ L, 0.95mmol) DMF (9mL) solution, and added on resin.Mixture is shaken at room temperature 2 hours.Repeat the mistake Journey twice, is carried out 1 hour with freshly prepared conjugate mixtures every time.Resin is washed into 10x with DMF (7mL), it is remaining free Amino 8mL 1/1/2 (v/v/v) Ac₂O/ pyridines/DMF end-blockings.

Carried out with DMF (47mL) solution of HOBt (0.68g, 5.03mmol), piperazine (3.00g, 34.83mmol) in 11c The cracking of Fmoc protection groups.Therefore, resin is incubated together with 10mL cleavage mixture 5 times, carries out 15min in rt every time. Resin is washed 7 times with DMF (7mL).

DIEA is added into 7 (266mg, 1.00mmol) and PyBOP (520mg, 1.00mmol) DMF (9mL) solution (209μL,1.20mmol).The mixture is added on resin, and shaken 2 hours at room temperature.Resin washs 7 with DMF (7mL) It is secondary, obtain resin 11e.By using the cleavage mixture 68.5/10/10/5/3.5/1 (v/w/v/v/v/ of 15mL precoolings (- 18 DEG C) V) TFA/DTT/ THIOANISOLEs/phenol/water/TIPS processing resins, realize peptide from the cracking of resin and the removal of protection group.Will be mixed Compound warms to room temperature, and stirs 3 hours.Thick 11f is precipitated in precooling (- 18 DEG C) ether and purified by RP-HPLC.Close And HPLC fractions be directly used in next step.

MS:M/z 1218.66=[M+4H]⁴⁺,([M+4H]⁴⁺Calculating single isotopic mass=1218.65)

In the HPLC product fractions (1L) of merging to 11f, 160mL 0.5M citrate buffer solutions (pH=5.00) are added With the solution in 9/1 (v/v) acetonitrile/water of the sulphur of 100mL 50mM 2,2 '-two double (pyridine-N-oxides).Mixture is existed Stir 4 hours, then diluted with the 1L water containing 0.1%TFA (v/v) at room temperature.11g is purified by RP-HPLC.

Yield:64.3mg (10.7 μm of ol, 6%) CNP-38- connectors-DMB*10TFA

MS:M/z 1218.15=[M+4H]⁴⁺,([M+4H]⁴⁺Calculating single isotopic mass=1218.14)

By cleavage mixture 100/5/3/2/1 (v/v/w/v/v) TFA/MSA/DTT/ for adding 45mL precoolings (- 18 DEG C) Water/THIOANISOLE then stirs 4 hours, to realize the cracking of Dmb protection groups to 11g (61.8mg, 10.3 μm of ol) at 0 DEG C. Thick 11h is precipitated in the ether of precooling (- 18 DEG C).Sediment is dissolved in 1/1 (v/v) second containing 0.1%TFA (v/v) In nitrile/aqueous solution, and it is incubated 4 hours at room temperature, to hydrolyze any TFA esters.11h is purified by RP-HPLC.

Yield:38.4mg (6.65 μm of ol, 65%) CNP-38- connectors-mercaptan * 10TFA

MS:M/z 1159.11=[M+4H]⁴⁺,([M+4H]⁴⁺Calculating single isotopic mass=1159.10)

PEG is added into 11h (34.6mg, the 5.99 μm of ol) solution in the 1mL water containing 0.1%TFA (v/v) 2x10kDa maleimides (GL2-200MA, 1.12g, 56.03 μm of ol of Sunbright) contain 0.1%TFA in 6.1mL (v/v) the solution in water, 0.5M lactic acid buffers (1.46mL, pH=4.00) are then added.Mixture is stirred into 4h in rt. Pass through RP-HPLC purified conjugation things 11i.

Yield:227mg (4.96 μm of ol, 83%) conjugate 11i*10HCl

Embodiment 12

Permanent Lys26CNP-38PEG4x10kDa conjugates 12g synthesis

According to following flow synthesis conjugate 12g：

To Fmoc-Lys (Fmoc)-OH (365mg, 0.62mmol) and PyBOP (322mg, 0.62mmol) DMF DIEA (0.11mL, 0.62mmol) is added in (4.6mL) solution.The mixture is added into resin 10a (2.0g, 0.21mmol). And suspension is shaken at room temperature 2 hours.Resin is washed 10 times with DMF (7mL).With HOBt (1.35g, 9.99mmol), DMF (94mL) solution of piperazine (6.00g, 69.66mmol) carries out the cracking of Fmoc protection groups in 12a.Therefore, by resin with splitting Solution mixture incubates 5 times together, carries out 15min in rt every time, obtains resin 12b.Then, resin is washed 7 with DMF (7mL) It is secondary.

Into 7 (283mg, 1.06mmol) and PyBOP (552mg, 1.06mmol) DMF (6.5mL) solution, DIEA is added (185 μ L, 1.06mmol), and add to resin 12b (2.07g, 0.10mmol/g, 0.21mmol).Mixture is shaken into 2h in rt. Then by resin DMF (7mL) and CH₂Cl₂(7mL) washs 10x, and is dried in a vacuum.

By using cleavage mixture 68.5/10/10/5/3.5/1 (v/w/v/v/v/v) TFA/ of 15mL precoolings (- 18 DEG C) DTT/ THIOANISOLEs/phenol/water/TIPS processing resins, realize peptide from the cracking of resin and the removal of protection group.By mixture temperature Heat stirs 2.5h to room temperature.Thick 12d is precipitated in precooling (- 18 DEG C) ether and purified by RP-HPLC.By merging HPLC fractions are directly used in next step.

MS:M/z 1172.37=[M+4H]⁴⁺,([M+4H]⁴⁺Calculating single isotopic mass=1172.37)

In the HPLC product fractions (390mL) of merging to 12d, 58.5mL 0.5M citrate buffer solutions (pH=is added 5.00) and 8.9mL 10mM 2,2 '-two sulphur double (pyridine-N-oxides) the solution in 1/1 (v/v) acetonitrile/water.Will be mixed 10min is stirred at room temperature in compound, is then diluted with the 400mL water containing 0.1%TFA (v/v).Purified by RP-HPLC 12e。

Yield:100mg (17.5 μm of ol, 8%, after 6 steps) CNP-38- connectors-Dmb*9TFA

MS:M/z 1171.87=[M+4H]⁴⁺,([M+4H]⁴⁺Calculating single isotopic mass=1171.86)

By cleavage mixture 100/5/3/2/1 (v/v/w/v/v) TFA/MSA/DTT/ for adding 65mL precoolings (- 18 DEG C) Water/THIOANISOLE then stirs 3.5h, to realize the cracking of Dmb protection groups to 12e (100mg, 17.5 μm of ol) at 0 DEG C.Will Thick 12f is precipitated in the ether of precooling (- 18 DEG C).Sediment is dissolved in the water containing 0.1%TFA (v/v), and in room temperature It is lower to be incubated 2 hours, to hydrolyze any TFA esters.12f is purified by RP-HPLC.

Yield:43.4mg (7.92 μm of ol, 45%) CNP-38- connectors-mercaptan * 9TFA

MS:M/z 1112.83=[M+4H]⁴⁺,([M+4H]⁴⁺Calculating single isotopic mass=1112.82)

PEG is added into 12f (39.6mg, the 7.22 μm of ol) solution in the 1mL water containing 0.1%TFA (v/v) 2x10kDa maleimides (GL2-200MA, 1.22g, 59.94 μm of ol of Sunbright) contain 0.1%TFA in 6.16mL (v/v) the solution in water, 0.5M lactic acid buffers (1.41mL, pH=4.20) are then added.Mixture is stirred into 4h in rt. Pass through RP-HPLC purified conjugation things 12g.

Yield:204mg (4.48 μm of ol, 57%) conjugate 12g*9HCl

Embodiment 13

PEG5kDa mercaptan 13c synthesis

According to following flow synthesis PEG5kDa mercaptan 13c:

To 13b (58.6mg, 0.15mmol), HOBt (22.9mg, 0.15mmol) and EDC hydrochlorides (28.8mg, In DCM (1.00mL) solution 0.15mmol), 2,4,6- trimethylpyridines (121mg, 1.00mmol) are added.Then first is added Epoxide PEG amine 5kDa 13a (500mg, 0.10mmol) DCM (4.00mL) solution, and mixture is stirred into 16h in rt.Evaporation Solvent, mixture is dissolved in ACN/ water, and is purified by RP-HPLC.The amount of solvent is reduced in a vacuum, and will be water-based residual Thing DCM (1x 100mL, 2x 50mL) is stayed to extract.The organic layer of merging is reduced to 20mL in a vacuum.Add TFA (1.6mL) and TES (3.5mL), and mixture is stirred into 4.5h in rt.13c is precipitated in ether, in -20 DEG C of storage over night, and It is dried in a vacuum.

Yield:372mg (72 μm of ol, 72%)

Embodiment 14

Permanent N- ends CNP-34PEG 5kDa conjugates 14e synthesis

According to following flow synthesis conjugate 14e:

CNP-34 14a (0.78g, 70 μ of side chain protected on the TCP tentagel resins with free N- ends Mol) the pre-swollen 30min in DMF.By maleimidocaproic acid (85.3mg, 0.40mmol), DIC (50.9mg, 0.40mmol) DMF (6mL) solution with Oxyma (57.4mL, 0.40mmol) is added on resin, and mixture is shaken in rt 30min.Then coupling is repeated once with freshly prepd conjugate solution.By resin DMF and CH₂Cl₂10x is washed, and is dried in vacuo Obtain 14b.

By using 6mL cleavage mixtures 100/3/2/1 (v/v/v/v) TFA/TES/ water/THIOANISOLE processing at room temperature Resin 1.5 hours, realize peptide from the cracking of resin and the removal of protection group.Thick peptide is precipitated in the ether of precooling (- 18 DEG C).

MS:M/z 937.77=[M+4H]⁴⁺,([M+4H]⁴⁺Calculating single isotopic mass=937.74)

Precipitation is dissolved in 15mL TFA.Add diphenyl sulfoxide (68.06mg, 0.34mmol) and methyl phenyl ethers anisole The 5mL TFA solution of (0.18mL, 1.68mmol).Trichloromethyl silane (0.47mL, 4.17mmol) is added, by mixture in room The lower stirring 15min of temperature.Ammonium fluoride (0.38g, 10.3mmol) is added, solution is stirred for 2min.By thick material in precooling (- 18 DEG C) ether in precipitate, and purified by RP-HPLC, obtain 14d.

Yield:8.30mg (1.78 μm of ol, 82% purity, 1.4%, after 3 steps) CNP-34-Malhx*8TFA

MS:M/z 937.26=[M+4H]⁴⁺,([M+4H]⁴⁺Calculating single isotopic mass=937.23)

To 14d (7.34mg, 1.57 μm of ol) in 200 μ L 1/1 (v/v) acetonitrile/waters containing 0.1%TFA (v/v) Solution in add 13c (20mg, 3.90 μm of ol) the solution in the 200 μ L water containing 0.1%TFA (v/v), then add 200 μ L 0.5M acetate buffers (pH=5.00).Mixture is incubated 30min at room temperature.Sewed by RP-HPLC purifying Compound 14e.

Yield:9.92mg (1.01 μm of ol, 57%) conjugate 14e*8TFA

Embodiment 15

Permanent N- ends CNP-38PEG 5kDa conjugates 15e synthesis

According to following flow synthesis conjugate 15e：

To synthesizing compound 15d as described in 14d, except that will have free N- ends on TCP tentagel resins The CNP-38 15a (1.34g, 0.12mmol) of side chain protected be used as initiation material.

Yield:15.6mg (2.94 μm of ol, 6.6%) CNP-38-Malhx*9TFA

MS:M/z 1064.05=[M+4H]⁴⁺,([M+4H]⁴⁺Calculating single isotopic mass=1064.04)

To synthesizing conjugate 15e as described in 14e, except 15d (8.34g, 1.58mmol) is used as into initiation material.

Yield:9.47mg (0.91 μm of ol, 31%) conjugate 15e*9TFA

Embodiment 16

Permanent Lys12CNP-34PEG 5kDa conjugates 16e synthesis

According to following flow synthesis conjugate 16e:

According to the program provided in material and method, by 1.00g (0.10mmol) have Boc protections N- ends and The CNP-34 of the side chain protected on TCP tentagel resins of the Lys12 side chains of ivDde protections carries out ivDde deprotections, Obtain 16a.

It is former except resin 16a (1.00g, 0.10mmol) is used as into starting to synthesizing compound 16d as described in 14d Material.

Yield:17.0mg (3.65 μm of ol, 3.7%) CNP-34-Lys12-Malhx*8TFA

MS:M/z 937.25=[M+4H]⁴⁺,([M+4H]⁴⁺Calculating single isotopic mass=937.23)

To synthesizing conjugate 16e as described in 14e, except 16d (17mg, 3.65 μm of ol) is used as into initiation material.

Yield:12.2mg (1.25 μm of ol, 34%) conjugate 16e*8TFA

Embodiment 17

Permanent Lys16CNP-34PEG 5kDa conjugates 17e synthesis

According to following flow synthesis conjugate 17e:

According to the program provided in material and method, by 0.78g (0.07mmol) have Boc protections N- ends and The CNP-34 of the side chain protected on TCP tentagel resins of the Lys16 side chains of ivDde protections carries out ivDde deprotections, Obtain 17a.

It is former except resin 17a (0.78g, 0.13mmol) is used as into starting to synthesizing compound 17d as described in 14d Material.

Yield:5.39mg (1.16 μm of ol, 1.7%) CNP-34-Lys16-Malhx*8TFA

To synthesizing conjugate 17e as described in 14e, except 17d (5.39mg, 1.16 μm of ol) is used as into initiation material.

Yield:10.7mg (1.09 μm of ol, 94%) conjugate 17e*8TFA

Embodiment 18

Permanent Lys22CNP-34PEG 5kDa conjugates 18e synthesis

According to following flow synthesis conjugate 18e:

According to the program provided in material and method, by 1.07g (0.11mmol) have Boc protections N- ends and The CNP-34 of the side chain protected on TCP tentagel resins of the Lys12 side chains of ivDde protections carries out ivDde deprotections, Obtain 18a.

It is former except resin 18a (1.07g, 0.11mmol) is used as into starting to synthesizing compound 18d as described in 14d Material.

Yield:5.20mg (1.12 μm of ol, 1.0%) CNP-34-Lys22-Malhx*8TFA

To synthesizing conjugate 18e as described in 14e, except 18d (5.2mg, 1.12 μm of ol) is used as into initiation material.

Yield:4.20mg (0.43 μm of ol, 38%) conjugate 18e*8TFA

Embodiment 19

Permanent Lys26CNP-38PEG 5kDa conjugates 19e synthesis

According to following flow synthesis conjugate 19e:

According to the program provided in material and method, by (0.865g, 0.10mmol) have Boc protections N- ends and The CNP-38 of the side chain protected on TCP tentagel resins of the Lys26 side chains of ivDde protections carries out ivDde deprotections, Obtain 19a.

It is former except resin 19a (0.865g, 0.10mmol) is used as into starting to synthesizing compound 19d as described in 14d Material.

Yield:10.3mg (1.95 μm of ol, 2.0%) CNP-38-Lys26-Malhx*9TFA

MS:M/z 1064.05=[M+4H]⁴⁺, (the single isotopic mass [M+4H] of calculating⁴⁺=1064.04)

To synthesizing conjugate 19e as described in 14e, except 19d (4.70mg, 1.10 μm of ol) is used as into initiation material.

Yield:3.20mg (0.31 μm of ol, 28%) conjugate 19e*9TFA

Embodiment 20

Vitro release kinetics

CNP conjugate 10f and 11i is dissolved in containing 3mM EDTA and 10mM methionines with about 1mg conjugates/mL concentration In the PBS of (pH 7.4).Solution is sterile filtered, and is incubated at 37 DEG C.Aliquot is taken out at time point and is led to Cross RP-HPLC and ESI-MS analyses.Map the UV- signal integration related to the CNP of release and relative to incubation time.

Application curves fitting software is estimated to discharge half-court accordingly.

As a result：

For conjugate 10f, acquisition 8.5d (± 1d) release half-life period.

For conjugate 11i, acquisition 9.5d (± 1.5d) release half-life period.

Embodiment 21

Neutral endopeptidase digests CNP variants in vitro

In order to determine in the presence of neutral endopeptidase (NEP) various CNP variants (including different peptide chain lengths and use The PEGylation of different pegylation sites and PEG molecules) vitro stability, establish NEP digestion determination method.The determination method With reference to t₀The reduction of the indigested CNP variants (being standardized with internal standard PFP) of-time point monitoring over time.

Specifically, by recombined human NEP (2.5 μ g/mL final concentrations) and standard Pentafluorophenol (PFP；40 μ g/mL final concentrations) The CNP variants (100 μ g CNP equivalents/mL) added in digestion buffer solution (50mM Tris-HCl, pH 7.4,10mM NaCl) In.Solution is incubated under 37 DEG C and 500rpm and is up to 4 days.Sample is taken at various time intervals.By adding TCEP ((three (2- carboxy ethyls) phosphines；25mM final concentrations) and mixture is incubated to 5min reduction and thermal denaturation under 95 DEG C, 500rpm Combination carrys out terminating reaction.Gained reaction product is determined using HPLC-MS.The half-life period of each CNP variants passes through CNP's and PFP The rate of change of HPLC-UV peak areas over time calculates.In order to compensate the change of proteinase activity, in the measurement of every batch CNP-38 or CNP-34 digestion is carried out as reference.

Table 1 is listed based on the different length of external NEP cracking measure and with the various PEG for being connected to different side chains The half-life period of the various CNP variants of molecule.

1) because the change of NEP catalytic activity between experiment, all CNP-34 half-life measurements form average value (4.15h), and coefficient of utilization by the half-life measurement value of CNP-34 conjugates with respect to the standardizing average values, to calculate adjustment T afterwards_1/2。

2) because the change of NEP catalytic activity between experiment, all CNP-38 half-life measurements form average value (12.10h), and coefficient of utilization is adjusted to relative standardizing average values of half-life measurement value of CNP-38 conjugates with calculating T after whole_1/2。

Resistance hierarchal order to NEP is as follows：The shorter CNP variants (CNP-34) of longer CNP- variants (CNP-38) More stable, CNP-34 is again more stable than shorter CNP-22.The order of PEG- connection sites is as follows：N- ends>It is close to ring>Ring.Cause This, for the conjugate tested, N-terminal PEG connections assign the highest stability of the proteolytic digestion for NEP. The CNP-38 of Lys26 PEGylation stability can increase with the increase of PEG sizes.

Embodiment 22

With CNP variants, feature cGMP is stimulated in NIH-3T3 cells

CNP variants are determined in the measure based on cell using NIH-3T3 cells (MEC system) Functional activity.These cells are in cell surface expression endogenous NPR-B.NPR-B is stimulated to cause the cGMP being commercially available to survey with CNP The intracellular for the second messenger cGMP that the method for determining detects produces.NIH-3T3 cells are being contained into 5%FBS and 5mM glutamine In 37 DEG C and 5%CO in DMEM F-12 culture mediums₂Lower cellar culture.For each measure, 50,000 cells are suspended again In stimulation buffer solution (Dulbecco ' the s PBS with IBMX), and it is incubated together with the CNP variants of various concentrations.CNP (is used 0.2%BSA dilutes in PBS).In 37 DEG C and 5%CO₂After lower incubation 30min, cell is cracked, and with commercially available cGMP TR-FRET measure (Cisbio, cGMP kit, catalog number (Cat.No.) 62GM2PEB) measure cGMP is horizontal.Pegylation CNP variants are total It is to be characterized in identical experiment batch compared with non-PEGylation version.If it is possible, the dose-response curve for passing through gained EC50- parameters (limited model with common slope) carry out the evaluation of residual activity.

Table 2：Become Pegylation CNP in the determination method based on cell of body measurement relative to non-Pegylation CNP to become The residual NPR-B activity of body

Compound	CNP variants	PEGylation	Residual activity [%]
				15e	CNP-38	5kDa PEG, N- ends	14
19e	CNP-38	5kDa PEG,Lys26	<1
				12g	CNP-38	4x10kDa PEG,Lys26	<<1

Compare the PEG connection sites of test, the connection at Lys26 (ring-lysine) place shows that highest functional activity drops It is low, and the connection of N- ends shows relatively high residual functional activity value.Increase PEG sizes cause CNP molecules preferably shielding and Lower residual functional activity.

Embodiment 23

After being treated 5 weeks with CNP-38 by daily subcutaneous bolus injection or continuous h inf, the increment study of FVB mouse

This research is carried out to test shadow of the CNP-38 daily continuous h infs of subcutaneous bolus injection vs. to growth of animal Ring.By daily subcutaneous bolus injection or by the continuous h inf in intrascapular region more than 35 days, to 21 days open countries to 22 day age Raw type FVB male mices (n=9/ groups) give 50nmol/kg/d CNP-38 or solvent (containing 5% sucrose and 1% benzylalcohol 30mM acetates pH 4).Continuous infusion is carried out the 1-2 weeks by the type of Alzet osmotic pumps 1002, is followed by 1004 types, continues 3-5 Week.In research the 7th day (type of pump 1002) or research the 25th day (type of pump 1004), the CNP- in pump is adjusted for average animal weight 38 concentration.By whole body linear measure longimetry and the X-ray measurement of right femur and shin bone, in measure growth in the 35th day.

The result for the animal treated by daily subcutaneous bolus injection：The 35th day, compared with the animal of vehicle treated, CNP-38 Total body length of the animal of processing is 110.2%, and right femur length is 105.6%, and right tibia length is 104.0%.

The result for the animal treated by continuous h inf：The 35th day, compared with the animal of vehicle treated, CNP-38 Total body length of the animal of processing is 121.7%, and right femur length is 107.5%, and right tibia length is 112.2%.

Conclusion be CNP-38 continuous h inf or related sustained release preparation (such as sustained release CNP-38 prodrugs) than daily skin Under inject more effectively trigger vertically and axial bone growth.

Embodiment 24

Pharmacokinetics of the permanent Lys26CNP-38PEG4x10kDa conjugates 12g in machin

This research is carried out to show 12g as the applicability of the model compound of sustained release CNP-38 prodrugs in machin. Male machin (2-4 year, 3.5-4.1kg) receives single intravenous injection (n=3 animals) or single SC (n=2 animals) 12g is applied, and dosage is 0.146mg CNP-38eq/kg.Blood sample is gathered after to be up to 168 hours, produces blood plasma.Such as material and Described in method, pass through the N- end tag peptide (sequences after quantitative Trypsin Induced：) and C- end tag peptide (sequences LQEHPNAR Row：IGSMSGLGC) plasma C NP-38 concentration is determined.

As a result：Dosage is applied and is well tolerated, without obvious uncomfortable sign during and after administration.Appoint in whole research When the reaction that is all not observed using site is waited.After intravenous injection, observed in 15min (the earliest time point of analysis) To CNP-38t_max, subsequent CNP-38 contents slow-decay, half-life period is about 24 hours.After hypodermic injection, CNP-38 concentration reaches Peak value, t_maxFor 48h.At 168 hours, CNP-38 concentration was still up to about 50%c_max.Bioavilability is about 50%.

After application until 168 hours, the similar PK profile of N- and C- end tag peptides is obtained, shows to deposit in conjugate In complete CNP-38.

CNP-38 stability represents permanent model compound in favourable lasting PK and conjugate within a couple of days Applicabilities of the Lys26CNP-38PEG 4x10kDa conjugates 12g after hypodermic injection as sustained release CNP-38 prodrugs.It can obtain Go out conclusion, the similar conjugate with the CNP-38 (such as 11i) of temporary transient Lys26 connections is that persistently level is provided within a couple of days Delivery of biologically active CNP-38 suitable CNP-38 prodrugs.

Embodiment 25

Pharmacokinetics of the temporary transient Lys26CNP-38PEG4x10kDa conjugates 11i in machin

It is the applicability in order to show 11i as sustained release CNP-38 prodrugs in machin to carry out this research.As implemented Studied described in example 24.Total CNP-38 contents (conjugated and release CNP-38) blood plasma water is analyzed as described in Example 24 It is flat.In order to analyze free CNP-38 plasma content, blood sample must be acidified after taking-up (such as by adding 20 volume % 0.5M sodium citrate buffer solutions pH 4) with stop from conjugate further release CNP-38.Free CNP-38 in blood plasma Level can be determined for example using the CNP antibody for combining CNP ring regions by ELISA, such as document (United States Patent (USP) 8,377,884B2) It is described, or determined by LC-MS/MS.

Embodiment 26

The temporary transient Lys26CNP-38PEG4x10kDa conjugates 11i pharmacodynamic study in machin

It is related to bone uptake to bone uptake using temporary transient Lys26CNP-38PEG4x10kDa conjugate 11i weekly treatments The influence of biomarker level is assessed in machin.Weekly 8 are subcutaneously injected with 16 or 56nmol/kg/ weeks normally Male juvenile machin (about 2 years old, study start when).Four monkeys were with the CNP-38 daily dosages of 8nmol/kg/ days It is subcutaneously injected, causes the intergal dose weekly of 56nmol/kg/ weeks.Other four monkeys apply solvent as control.That treats is total Length is 6 months.Growth plate expansion and bone are carried out by digital X-ray and magnetic resonance imaging and externally measured limbs and body length The various measurements of growth.Periodic collection blood and urine specimen carry out clinicopathologia and measurement.At the end of research, carry out big Body pathology, tissue samples are histologically assessed to assess efficacy and saferry.

Abbreviation:

ACH achondroplasias

ACN acetonitriles

AcOH acetic acid

Bn benzyls

Boc tertbutyloxycarbonyls

BSA bovine serum albumin(BSA)s

CGMP cyclic guanylic acid monophosphates

CNP c-type natriuretic peptides

COMU (1- cyano group -2- ethyoxyl -2- oxo ethyleneiminos epoxide) dimethylamino

Base-morpholino-carbonium ion hexafluorophosphate

Conc. it is dense

D days

DBU 1,3- diazabicylos [5.4.0] endecatylene

DCC N, N '-dicyclohexylcarbodiimide

DCM dichloromethane

DIC N, N '-DIC

DIEA N, N- diisopropyl ethyl amines

DIPEA N, N- diisopropyl ethyl amines

DMAP dimethylaminos-pyridine

The Eagle culture mediums of DMEM Dulbecco improvement

Dmb 2,4- dimethyl benzyls

The eagle culture mediums of DMEM Dulbecco improvement

DMF N,N-dimethylformamides

DMSO dimethyl sulfoxide (DMSO)s

DTT dithiothreitol (DTT)s

EC50 half maximum valid density

EDC 1- ethyls -3- (3- dimethylaminopropyls) carbodiimide

EDTA ethylenediamine tetra-acetic acids

ELISA enzyme linked immunosorbent assay (ELISA)s

Eq equivalent stoichiometric equivalents

ESI-MS LC-MS spectrometry methods

Et ethyls

EtOAc ethyl acetate

EtOH ethanol

FBS hyclones

FGFR3 fibroblast growth factor receptor3s

Fmoc 9- fluorenylmethyloxycarbonyls

H hours

HATU O- (7- azepine benzos triazol-1-yl)-N, N, N ', N '-tetramethylureaSix

Fluorophosphate

HCH osteochondrodysplasias

HFIP hexafluoroisopropanols

HPLC high performance liquid chromatography

HOBt N- hydroxybenzotriazoles

IBMX 3-isobutyl-1-methylxanthines

IPrOH 2- propyl alcohol

Iv is intravenous

IvDde 4,4- dimethyl -2,6- dioxocyclohexyl -1- subunits) -3- methyl butyls

LC liquid chromatograies

LTQ linear trap quadrupole rods

Mal 3- Maleimido propyl group

Me methyl

MeOH methanol

Min minutes

Mmt monomethoxytrityls

MS mass spectrums/mass spectrography

MSA methanesulfonic acids

MW molecular weight

M/z mass-to-charge ratioes

NEP neutral endopeptidases

NHS n-hydroxysuccinimides

NPR natriuretic peptide receptors

OtBu tert-butyl group epoxides

PBS phosphate buffered saline (PBS)s

PEG PEGs

PFP Pentafluorophenols

PH hydrogen ion concentrations (potentia Hydrogenii)

Pr propyl group

PyBOP BTAs -1- bases-epoxide tripyrrole alkane Ji Phosphonium hexafluorophosphates

The Q-TOF quadrupole rod flight time

RP-HPLC RPLCs

Rpm revolutions per minutes

Rt room temperatures

SIM single ion monitorings

SEC size exclusion chromatographies

Sc is subcutaneous

t_1/2Half-life period

TCEP tri- (2- carboxy ethyls) phosphine

TCP trityl chloride polystyrenes

TD thanatophoric dysplasias

TES triethyl silicanes

TFA trifluoroacetic acids

THF tetrahydrofurans

TIPS tri isopropyl silanes

TMEDA N, N, N ' N '-tetramethylethylenediamine

Tmob 2,4,6- trimethoxy benzyls

TR-FRET time-resolved fluorescence energy transfer (TR-FRET)s

Trt trityl groups, trityl

UPLC ultra performance liquid chromatographies

UV ultraviolets

Vs. compare

The mono- quadrupole rods of ZQ

Claims

1.CNP prodrugs or its pharmaceutically acceptable salt, wherein the prodrug has formula (Ia) or (Ib)

Wherein

- D is CNP parts；

-L¹- it is reversible prodrug connector part；

-L²- it is single chemical bond or interval body portion；

- Z is water-solubility carrier part；

Y is to be selected from following integer：1st, 2,3,4 and 5.

2.CNP prodrugs or its pharmaceutically acceptable salt, it includes conjugate D-L, wherein

- D is CNP parts；And

- L includes reversible prodrug connector part-L¹-；

-L²- it is single chemical bond or interval body portion；And

- Z ' is water insoluble carrier part.

3. the CNP prodrugs or its pharmaceutically acceptable salt of claim 2, wherein-Z ' it is hydrogel.

4. the CNP prodrugs or its pharmaceutically acceptable salt of claim 1, wherein the CNP prodrugs have formula (Ia).

5. the CNP prodrugs or its pharmaceutically acceptable salt of claim 1 or 4, wherein x is 1.

6. the CNP prodrugs or its pharmaceutically acceptable salt of any one in claim 1 to 5, wherein CNP parts have SEQ ID NO:25 or SEQ ID NO:24 sequence.

7. the CNP prodrugs or its pharmaceutically acceptable salt of any one in claim 1 to 6, wherein CNP parts have SEQ ID NO:24 sequence.

8. the CNP prodrugs or its pharmaceutically acceptable salt of any one in claim 1 to 7, wherein-L¹- it is conjugated to-D ring The side chain of partial amino acid residue or the extremely main chain of-D loop section.

9. the CNP prodrugs or its pharmaceutically acceptable salt of any one in claim 1 to 8, wherein-L¹- it is conjugated to-D ring The side chain of partial amino acid residue, the amino acid are selected from：Histidine, lysine, tryptophan, serine, threonine, junket ammonia Acid, aspartic acid, glutamic acid and arginine.

10. the CNP prodrugs or its pharmaceutically acceptable salt of any one in claim 1 to 9, wherein-D has SEQ ID NO:24 sequence, and-L¹- it is conjugated to lysine in-D position 26.

11. the CNP prodrugs or its pharmaceutically acceptable salt of any one, which part-L in claim 1 to 10¹- there is formula (II):

- X- is-C (R⁴R^4a)-；-N(R⁴)-；-O-；-C(R⁴R^4a)-C(R⁵R^5a)-；-C(R⁵R^5a)-C(R⁴R^4a)-；-C(R⁴R^4a)-N (R⁶)-；-N(R⁶)-C(R⁴R^4a)-；-C(R⁴R^4a)-O-；-O-C(R⁴R^4a)-；Or-C (R⁷R^7a)-；

X¹It is C；Or S (O)；

-X²- it is-C (R⁸R^8a)-；Or-C (R⁸R^8a)-C(R⁹R^9a)-；

=X³It is=O；=S；Or=N-CN；

-R¹、-R^1a、-R²、-R^2a、-R⁴、-R^4a、-R⁵、-R^5a、-R⁶、-R⁸、-R^8a、-R⁹、-R^9aIndependently selected from-H；And C_1-6Alkyl；

-R³、-R^3aIndependently selected from-H；And C_1-6Alkyl, if on condition that-R³、-R^3aIn one or two be not -H, then it Pass through SP³The carbon atom of-hydridization is connected on the N that they are connected；

-R⁷It is-N (R¹⁰R^10a)；Or-NR¹⁰- (C=O)-R¹¹；

Optionally, one or more-R^1a/-R^4a、-R^1a/-R^5a、-R^1a/-R^7a、-R^4a/-R^5a、-R^8a/-R^9aTo forming chemical bond；

Optionally, one or more-R¹/-R^1a、-R²/-R^2a、-R⁴/-R^4a、-R⁵/-R^5a、-R⁸/-R^8a、-R⁹/-R^9aPair and they The atom connected forms C together_3-10Cycloalkyl；Or 3- is to 10- circle heterocycles bases；

Optionally, one or more-R¹/-R⁴、-R¹/-R⁵、-R¹/-R⁶、-R¹/-R^7a、-R⁴/-R⁵、-R⁴/-R⁶、-R⁸/-R⁹、-R²/- R³Pair ring A is formed together with the atom that they are connected；

A is selected from phenyl；Naphthyl；Indenyl；Indanyl；Tetrahydro naphthyl；C_3-10Cycloalkyl；3- is to 10- circle heterocycles bases；With 8- to 11- 9-membered heterobicyclic base；And

Wherein-L¹- by-L²- Z or-L²- Z ' substitutes, and wherein-L¹- be optionally further substituted, on condition that the use in formula (II) The hydrogen of asterisk mark is not by-L²- Z or-L²- Z ' or substituent are replaced；

Wherein

-L²- it is single chemical bond or interval body；

- Z is water-solubility carrier；And

- Z ' is water insoluble carrier.

12. the CNP prodrugs or its pharmaceutically acceptable salt of claim 11, wherein-X- is-C (R⁴R^4a)-or-N (R⁴)-。

13. the CNP prodrugs or its pharmaceutically acceptable salt of claim 11 or 12, wherein-R⁴By-L²- Z or-L²- Z ' substitutes.

14. the CNP prodrugs or its pharmaceutically acceptable salt of any one, wherein X in claim 11 to 13¹It is C.

15. the CNP prodrugs or its pharmaceutically acceptable salt of any one in claim 11 to 14, wherein=X³It is=O.

16. the CNP prodrugs or its pharmaceutically acceptable salt of any one in claim 11 to 15, wherein-X²- it is-C (R⁸R^8a)-。

17. the CNP prodrugs or its pharmaceutically acceptable salt of any one in claim 11 to 16, wherein-R¹With-R^1aBe- H。

18. the CNP prodrugs or its pharmaceutically acceptable salt of any one in claim 11 to 17, wherein-R²With-R^2aBe- H。

19. the CNP prodrugs or its pharmaceutically acceptable salt of any one in claim 11 to 18, wherein-R³It is-H, and- R^3aIt is methyl.

20. the CNP prodrugs or its pharmaceutically acceptable salt of any one in claim 11 to 19, wherein-R⁴With-R^4aBe- H。

21. the CNP prodrugs or its pharmaceutically acceptable salt of any one in claim 11 to 20, wherein-R⁸With-R^8aBe- H。

22. the CNP prodrugs or its pharmaceutically acceptable salt of any one in claim 1 to 21, wherein-L²- it is selected from-T- ,-C (O)O-、-O-、-C(O)-、-C(O)N(R^y1)-、-S(O)₂N(R^y1)-、-S(O)N(R^y1)-、-S(O)₂-、-S(O)-；-N(R^y1)S (O)₂N(R^y1a)-、-S-、-N(R^y1)-、-OC(OR^y1)(R^y1a)-、-N(R^y1)C(O)N(R^y1a)-、-OC(O)N(R^y1)-、C_1-50Alkane Base, C_2-50Alkenyl and C_2-50Alkynyl；Wherein-T-, C_1-50Alkyl, C_2-50Alkenyl and C_2-50Alkynyl optionally by one or more identical or Different-R^y2Substitution, and wherein C_1-50Alkyl, C_2-50Alkenyl and C_2-50Alkynyl is optionally selected from following base by one or more Group interrupts：-T-、-C(O)O-、-O-、-C(O)-、-C(O)N(R^y3)-、-S(O)₂N(R^y3)-、-S(O)N(R^y3)-、-S(O)₂-、-S (O)-、-N(R^y3)S(O)₂N(R^y3a)-、-S-、-N(R^y3)-、-OC(OR^y3)(R^y3a)-、-N(R^y3)C(O)N(R^y3a)-and-OC (O) N(R^y3)-；

-R^y1With-R^y1aIt is independently selected from-H ,-T, C_1-50Alkyl, C_2-50Alkenyl and C_2-50Alkynyl；Wherein-T, C_1-50Alkyl, C_2-50Alkenyl and C_2-50Alkynyl is optionally by one or more identical or different-R^y2Substitution, and wherein C_1-50Alkyl, C_2-50Alkene Base and C_2-50Alkynyl is optionally interrupted by one or more selected from following group：-T-、-C(O)O-、-O-、-C(O)-、-C(O)N (R^y4)-、-S(O)₂N(R^y4)-、-S(O)N(R^y4)-、-S(O)₂-、-S(O)-、-N(R^y4)S(O)₂N(R^y4a)-、-S-、-N (R^y4)-、-OC(OR^y4)(R^y4a)-、-N(R^y4)C(O)N(R^y4a)-and-OC (O) N (R^y4)-；

Each T independently selected from：Phenyl, naphthyl, indenyl, indanyl, tetrahydro naphthyl, C_3-10Cycloalkyl, 3- to 10- circle heterocycles base, 8- is to 11- 9-membered heterobicyclics base, 8- to the more ring groups of 30- member carbon and the miscellaneous more ring groups of 8- to 30- members；Wherein each T is independently optionally by one Individual or multiple identical or different-R^y2Substitution；

Respectively-R^y2Independently selected from：Halogen ,-CN, oxo (=O) ,-COOR^y5、-OR^y5、-C(O)R^y5、-C(O)N(R^y5R^y5a)、-S (O)₂N(R^y5R^y5a)、-S(O)N(R^y5R^y5a)、-S(O)₂R^y5、-S(O)R^y5、-N(R^y5)S(O)₂N(R^y5aR^y5b)、-SR^y5、-N (R^y5R^y5a)、-NO₂、-OC(O)R^y5、-N(R^y5)C(O)R^y5a、-N(R^y5)S(O)₂R^y5a、-N(R^y5)S(O)R^y5a、-N(R^y5)C(O) OR^y5a、-N(R^y5)C(O)N(R^y5aR^y5b)、-OC(O)N(R^y5R^y5a) and C_1-6Alkyl；Wherein C_1-6Alkyl is optionally one or more Identical or different halogen substitution；And

Respectively-R^y3、-R^y3a、-R^y4、-R^y4a、-R^y5、-R^y5aWith-R^y5bIndependently selected from：- H and C_1-6Alkyl, wherein C_1-6Alkyl is optional Substituted by one or more identical or different halogens.

23. the CNP prodrugs or its pharmaceutically acceptable salt of any one in claim 1 to 22, wherein-L²- it is C_1-20Alkyl Chain, it is optionally independently selected from following group by one or more and interrupted：- O- ,-T- and-C (O) N (R^y1)-；And the C_1-20Alkyl Chain is optionally independently selected from following group by one or more and substituted：- OH ,-T and-C (O) N (R^y6R^y6a)；Wherein-R^y1、-R^y6、- R^y6aIndependently selected from H and C_1-4Alkyl, and wherein T is selected from phenyl, naphthyl, indenyl, indanyl, tetrahydro naphthyl, C_3-10Cycloalkanes Base, 3- to 10- circle heterocycles base, 8- to 11- 9-membered heterobicyclics base, 8- to the more ring groups of 30- member carbon and the miscellaneous more ring groups of 8- to 30- members.

24. the CNP prodrugs or its pharmaceutically acceptable salt of any one in claim 1 to 23, wherein-L²- there is formula (i)

Wherein

The dotted line instruction marked with asterisk is connected to-L¹-；

Unlabelled dotted line instruction is connected to-Z or-Z '；

-R¹Selected from-H, C_1-6Alkyl, C_2-6Alkenyl and C_2-6Alkynyl；

N is selected from 0,1,2,3,4,5,6,7,8,9,10,11,12,13,14,15,16,17 and 18；And

The part of wherein formula (i) is optionally further substituted.

25. claim 1 or 4 is to the prodrug or its pharmaceutically acceptable salt of any one in 24, wherein-Z have 5 to 200kDa molecular weight.

26. claim 1 or 4 includes C to the prodrug or its pharmaceutically acceptable salt of any one in 25, wherein carrier-Z_8-24 Alkyl or polymer.

27. claim 1 or 4 is to the prodrug or its pharmaceutically acceptable salt of any one in 26, wherein-Z include be selected from Under polymer：2- methylacryloyls-epoxide ethylphosphocholine, poly- (acrylic acid), poly- (acrylate), poly- (acryloyl Amine), poly- (alkoxy) polymer, poly- (acid amides), poly- (amidoamines), poly- (amino acid), poly- (acid anhydrides), poly- (asparagine), Poly- (butyric acid), poly- (glycolic), polybutylene terephthalate, poly- (caprolactone), poly- (carbonic ester), poly- (alpha-cyanoacrylate Ester), poly- (DMAA), poly- (ester), poly- (ethene), PEG, poly- (oxirane), poly- (etherophosphoric acid), Poly- (ethylOxazoline), poly- (glycolic), poly- (dihydroxypropyl ethyl ester), it is poly- (hydroxyethyl-Oxazoline), poly- (hydroxyl first Base acrylate), poly- (hydroxypropylmethacrylamide), poly- (hydroxy propyl methacrylate), poly- (hydroxypropylAzoles Quinoline), poly- (iminocarbonic ester), poly- (lactic acid), polylactic-co-glycolic acid, poly- (Methacrylamide), poly- (methyl-prop Olefin(e) acid ester), poly- (methylOxazoline), poly- (organic phosphine nitrile), poly- (ortho esters), it is poly- (Oxazoline), poly- (propane diols), poly- (silica Alkane), poly- (carbamate), poly- (vinyl alcohol), poly- (vinyl amine), poly- (vinyl methyl ether), poly- (ethenyl pyrrolidone Ketone), silicone, cellulose, carboxymethyl cellulose, HYDROXY PROPYL METHYLCELLULOSE, chitin, chitosan, glucan, dextrin, gelatin, Hyaluronic acid and derivative, the hyaluronic acid of functionalization, mannosan, pectin, rhamnose galacturonic acid glycan, starch, hydroxyl Base alkyl starches, hydroxy ethyl starch and other polymer based on carbohydrate, xylan and its copolymers.

28. claim 1 or 4 is gathered to the CNP prodrugs or its pharmaceutically acceptable salt of any one in 27 wherein-Z is side chain Compound.

29. claim 1 or 4 is to the CNP prodrugs or its pharmaceutically acceptable salt of any one in 28, wherein-Z has at least 10kDa molecular weight.

30. the CNP prodrugs or its pharmaceutically acceptable salt of any one in claim 1 to 29, wherein-Z or-Z' includes portion Point

31. claim 1 or 4 is to the CNP prodrugs or its pharmaceutically acceptable salt of any one in 30, wherein-Z includes formula (a) part

Wherein

Dotted line instruction is connected to-L²- or to-Z remainder；

BP^aIt is to be selected from following branch point：-N<、-CR<With>C<；

- R is selected from-H and C_1-6Alkyl；

If BP^aIt is-N<Or-CR<, then a is 0, and if BP^aIt is>C<, then n is 1；

-S^a-、-S^a’-、-S^a”- and-S^a”’- it is chemical bond or selected from C independently of each other_1-50Alkyl, C_2-50Alkenyl and C_2-50Alkynyl； Wherein C_1-50Alkyl, C_2-50Alkenyl and C_2-50Alkynyl is optionally by one or more identical or different-R¹Substitution, and wherein C_1-50 Alkyl, C_2-50Alkenyl and C_2-50Alkynyl is optionally interrupted by one or more selected from following group：-T-、-C(O)O-、-O-、-C (O)-、-C(O)N(R²)-、-S(O)₂N(R²)-、-S(O)N(R²)-、-S(O)₂-、-S(O)-、-N(R²)S(O)₂N(R^2a)-、- S-、-N(R²)-、-OC(OR²)(R^2a)-、-N(R²)C(O)N(R^2a)-and-OC (O) N (R²)-；

Respectively-T- independently selected from：Phenyl, naphthyl, indenyl, indanyl, tetrahydro naphthyl, C_3-10Cycloalkyl, 3- to 10- circle heterocycles Base, 8- to 11- 9-membered heterobicyclics base, 8- to the more ring groups of 30- member carbon and the miscellaneous more ring groups of 8- to 30- members；Wherein each-T- is independently optional By one or more identical or different-R¹Substitution；

Respectively-R¹Independently selected from：Halogen ,-CN, oxo (=O) ,-COOR³、-OR³、-C(O)R³、-C(O)N(R³R^3a)、-S(O)₂N (R³R^3a)、-S(O)N(R³R^3a)、-S(O)₂R³、-S(O)R³、-N(R³)S(O)₂N(R^3aR^3b)、-SR³、-N(R³R^3a)、-NO₂、-OC (O)R³、-N(R³)C(O)R^3a、-N(R³)S(O)₂R^3a、-N(R³)S(O)R^3a、-N(R³)C(O)OR^3a、-N(R³)C(O)N (R^3aR^3b)、-OC(O)N(R³R^3a) and C_1-6Alkyl；Wherein C_1-6Alkyl is optionally taken by one or more identical or different halogens Generation；

Respectively-R²、-R^2a、-R³、-R^3aWith-R^3bIndependently selected from：- H and C_1-6Alkyl, wherein C_1-6Alkyl is optionally one or more Identical or different halogen substitution；And

-P^a’、-P^a”With-P^a”’It is independently polymer moieties.

32. claim 1 or 4 is to the CNP prodrugs or its pharmaceutically acceptable salt of any one in 31, wherein-Z has formula (d)

Wherein

Dotted line instruction is connected to-L²-；

-Z^b- it is selected from C_1-50Alkyl, C_2-50Alkenyl and C_2-50Alkynyl；Wherein C_1-50Alkyl, C_2-50Alkenyl and C_2-50Alkynyl is optionally by one Individual or multiple identical or different-R¹Substitution and wherein C_1-50Alkyl, C_2-50Alkenyl and C_2-50Alkynyl is optionally one or more Interrupted selected from following group：-T-、-C(O)O-、-O-、-C(O)-、-C(O)N(R²)-、-S(O)₂N(R²)-、-S(O)N (R²)-、-S(O)₂-、-S(O)-、-N(R²)S(O)₂N(R^2a)-、-S-、-N(R²)-、-OC(OR²)(R^2a)-、-N(R²)C(O)N (R^2a)-and-OC (O) N (R²)-；

Respectively-R²、-R^2a、-R³、-R^3aWith-R^3bIndependently selected from：- H and C_1-6Alkyl, wherein C_1-6Alkyl is optionally one or more Identical or different halogen substitution；

And

-Z^aIt is

Wherein

BP^a、-S^a-、-S^a’-、-S^a”-、-S^a”’-、-P^a’、-P^a”、-P^a”’With a as defined in claim 31 use.

33. claim 1 or 4 is to the CNP prodrugs or its pharmaceutically acceptable salt of any one in 32, wherein-Z has formula (e)

Wherein

Dotted line instruction is connected to-L²-；

E is selected from 1,2,3,4,5,6,7,8,9,10,11,12,13,14 and 15；And

-Z^aIt is

Wherein

B1 is selected from 0,1,2,3,4,5,6,7 and 8；

B2 is selected from 1,2,3,4,5,6,7 and 8；

B3 is the integer in 150 to 1000 scopes and including end points；And

B4 is the integer in 150 to 1000 scopes and including end points.

34. the CNP prodrugs or its pharmaceutically acceptable salt of claim 33, wherein e is that 5, b1 is that 2, b2 is 3, and b3 and b4 All it is about 450.

35. claim 1 or 4 is to the CNP prodrugs or its pharmaceutically acceptable salt of any one in 31, wherein-Z has formula (f)

Wherein

Dotted line instruction is connected to-L²-；

BP^fIt is to be selected from following branch point：-N<、-CR<With>C<；

- R is selected from-H and C_1-6Alkyl；

If BP^fIt is-N<Or-CR<, f is 0, and if BP^fIt is>C<, f is 1；

-S^f-、-S^f’-、-S^f”- and-S^f”’- it is independently chemical bond or independently selected from C_1-50Alkyl, C_2-50Alkenyl and C_2-50Alkynes Base；Wherein C_1-50Alkyl, C_2-50Alkenyl and C_2-50Alkynyl is optionally by one or more identical or different-R¹Substitute and wherein C_1-50Alkyl, C_2-50Alkenyl and C_2-50Alkynyl is optionally interrupted by one or more selected from following group：-T-、-C(O)O-、- O-、-C(O)-、-C(O)N(R²)-、-S(O)₂N(R²)-、-S(O)N(R²)-、-S(O)₂-、-S(O)-、-N(R²)S(O)₂N (R^2a)-、-S-、-N(R²)-、-OC(OR²)(R^2a)-、-N(R²)C(O)N(R^2a)-and-OC (O) N (R²)-；

Each R¹Independently selected from：Halogen ,-CN, oxo (=O) ,-COOR³、-OR³、-C(O)R³、-C(O)N(R³R^3a)、-S(O)₂N (R³R^3a)、-S(O)N(R³R^3a)、-S(O)₂R³、-S(O)R³、-N(R³)S(O)₂N(R^3aR^3b)、-SR³、-N(R³R^3a)、-NO₂、-OC (O)R³、-N(R³)C(O)R^3a、-N(R³)S(O)₂R^3a、-N(R³)S(O)R^3a、-N(R³)C(O)OR^3a、-N(R³)C(O)N (R^3aR^3b)、-OC(O)N(R³R^3a) and C_1-6Alkyl；Wherein C_1-6Alkyl is optionally taken by one or more identical or different halogens Generation；

And

-Z^a’、-Z^a‘’With-Z^a”’It is independently

Wherein

36. the CNP prodrugs or its pharmaceutically acceptable salt of any one in claim 1,4 to 31 or 35, wherein-Z has Formula (g)

Wherein

Dotted line instruction is connected to-L²-；

-S^g-、-S^g’- and-S^g”- independently selected from C_1-50Alkyl, C_2-50Alkenyl and C_2-50Alkynyl；Wherein C_1-50Alkyl, C_2-50Alkenyl And C_2-50Alkynyl is optionally by one or more identical or different-R¹Substitution and wherein C_1-50Alkyl, C_2-50Alkenyl and C_2-50Alkynes Base is optionally interrupted by one or more selected from following group：-T-、-C(O)O-、-O-、-C(O)-、-C(O)N(R²)-、-S(O)₂N(R²)-、-S(O)N(R²)-、-S(O)₂-、-S(O)-、-N(R²)S(O)₂N(R^2a)-、-S-、-N(R²)-、-OC(OR²) (R^2a)-、-N(R²)C(O)N(R^2a)-and-OC (O) N (R²)-；

And

-Z^aWith-Z^a’It is independently

Wherein

37. the CNP prodrugs or its pharmaceutically acceptable salt of any one in claim 1,4 to 31,35 or 36, wherein-Z has There is formula (h)

Wherein

Dotted line instruction is connected to-L²-；And

Respectively-Z^cIt is part

Wherein

Each c1 is independently the integer in about 200 to 250 scopes.

38. claim 1 or 4 is to the CNP prodrugs or its pharmaceutically acceptable salt of any one in 34, wherein before the CNP Medical instrument has formula (IIe)

Wherein

Unlabelled dotted line instruction is by forming amido link connection most on the-D of CNP parts nitrogen；

And

The dotted line instruction marked with asterisk is connected to part

Wherein

Each c1 is independently the integer in 400 to 500 scopes.

39. the CNP prodrugs or its pharmaceutically acceptable salt of any one in claim 1,4 to 34 or 38, wherein the CNP Prodrug has formula (IIe ')

Wherein

Unlabelled dotted line instruction is connected to SEQ ID NO by forming amido link:The bad ammonia in position 26 of 24 CNP parts The nitrogen that the side chain of acid provides；And

The dotted line instruction marked with asterisk is connected to part

Wherein

Each c1 is independently the integer in 400 to 500 scopes.

40. the CNP prodrugs or its pharmaceutically acceptable salt of any one in claim 1,4 to 34, wherein the CNP prodrugs With formula (IIe-i ')

Wherein

The dotted line instruction marked with asterisk is connected to part

Wherein

Each c1 is independently the integer in 400 to 500 scopes.

41. the CNP prodrugs or its pharmaceutically acceptable salt of any one in claim 1,4 to 34, wherein the CNP prodrugs With formula (IIe-ii ')

Wherein

The dotted line instruction marked with asterisk is connected to part

Wherein

Each c1 is independently the integer in 400 to 500 scopes.

42. the CNP prodrugs or its pharmaceutically acceptable salt of any one, wherein institute in claim 1,4 to 31 or 35 to 37 Stating CNP prodrugs has formula (IIf)

Wherein

And

Wherein

Respectively-Z^aIt is

Wherein

Each c1 is independently the integer in 200 to 250 scopes.

43. the CNP prodrugs or its pharmaceutically acceptable salt of any one in claim 1,4 to 31,35 to 37 or 42, wherein The CNP prodrugs have formula (IIf ')

Wherein

Each Z^aIt is

Wherein

Each c1 is independently the integer in 200 to 250 scopes.

44. the CNP prodrugs or its pharmaceutically acceptable salt of any one, wherein institute in claim 1,4 to 31 or 35 to 37 Stating CNP prodrugs has formula (IIf-i ')

Wherein

Each Z^aIt is

Wherein

Each c1 is independently the integer in 200 to 250 scopes.

45. the CNP prodrugs or its pharmaceutically acceptable salt of any one, wherein institute in claim 1,4 to 31 or 35 to 37 Stating CNP prodrugs has formula (IIf-ii ')

Wherein

Each Z^aIt is

Wherein

Each c1 is independently the integer in 200 to 250 scopes.

46. the prodrug of any one or its pharmaceutically acceptable salt in Claims 1-4 5, wherein the CNP prodrugs is residual Activity is stayed to be less than 10%.

47. pharmaceutical composition, it includes in Claims 1-4 6 at least one CNP prodrugs of any one or it pharmaceutically may be used The salt of receiving and at least one excipient.

48. the pharmaceutical composition of claim 47, wherein described pharmaceutical composition have pH 4 to a pH 6 pH scopes, and including End points.

49. the prodrug of any one or its pharmaceutically acceptable salt or the medicine of claim 47 or 48 in Claims 1-4 6 Purposes of the compositions as medicine.

50. the prodrug of any one or its pharmaceutically acceptable salt or the medicine of claim 47 or 48 in Claims 1-4 6 Purposes of the compositions in the treatment method of the disease of available CNP treatments.

51. the purposes of claim 50, wherein the disease is selected from achondroplasia, osteochondrodysplasia, of short and small stature, dwarf It is scholar disease, osteochondrodysplasia, thanatophoric dysplasia, osteogenesis imperfecta, achondrogenesis, punctatae, pure Zygote achondroplasia, camptomelic dysplasia, congenital lethal hypophosphatasia, perinatal period lethal type skeletonization are not Entirely, short rib refers to syndrome, limb root type punctatae, Jansen type metaphyseal dysplasia, congenital centrum epiphysis more Dysplasia, Atelosteogenesis, diastrophic dysplasia, congenital short femur, Lange type mesomelic dysplasia, It is Nievergelt types mesomelic dysplasia, Robinow syndromes, Reinhardt syndromes, acrodysostosis, peripheral Dysostosis, Kniest depauperations, fibrocartilage generation, Roberts syndromes, acra mesomelic dysplasia, manomelia Disease, Morquio syndromes, Kniest syndromes, metatrophic dysplasia, vertebra epiphysis metaphyseal dysplasia, nerve Fibromatosis, Legius syndromes, LEOPARD syndromes, Noonan syndromes, hereditary gingival fibromatosis, 1 type nerve are fine Tie up knurl disease, Legius syndromes, Cardiofaciocutaneous syndromes, Costello syndromes, SHOX deficiency diseases, special hair Property of short and small stature, growth hormone deficiency, osteoarthritis, cleidocranial dysostosis, craniosynostosis (such as Muenke synthesis Sign, Crouzon syndromes, Apert syndromes, Jackson-Weiss syndromes, Pfeiffer syndromes or Crouzonodermoskeletal syndromes), refer to/toe type, brachydactylia/toe, camptodactylia/toe, polydactyly, zygodactyly, Dyssegmental depauperations, enchondrosis, fibrous dysplasia, hereditary multiple exostoses, hypophosphate Hypophosphatemic rickets, Jaffe-Lichtenstein syndromes, Marfan syndromes, McCune-Albright syndromes, bone are hard Change disease and osteopoikilosis.

52. the purposes of claim 50, wherein the disease is ophthalmology disease.

53. the purposes of claim 50 or 51, wherein the disease is achondroplasia.

54. the CNP prodrugs of any one or its pharmaceutically acceptable salt or claim 47 or 48 in Claims 1-4 6 Pharmaceutical composition, which is used to prepare to be used to treat, can use the purposes in the medicine of the CNP diseases treated.

55. the purposes of claim 54, wherein the disease is selected from achondroplasia, osteochondrodysplasia, of short and small stature, dwarf It is scholar disease, osteochondrodysplasia, thanatophoric dysplasia, osteogenesis imperfecta, achondrogenesis, punctatae, pure Zygote achondroplasia, camptomelic dysplasia, congenital lethal hypophosphatasia, perinatal period lethal type skeletonization are not Entirely, short rib refers to syndrome, limb root type punctatae, Jansen type metaphyseal dysplasia, congenital centrum epiphysis more Dysplasia, Atelosteogenesis, diastrophic dysplasia, congenital short femur, Lange type mesomelic dysplasia, It is Nievergelt types mesomelic dysplasia, Robinow syndromes, Reinhardt syndromes, acrodysostosis, peripheral Dysostosis, Kniest depauperations, fibrocartilage generation, Roberts syndromes, acra mesomelic dysplasia, manomelia Disease, Morquio syndromes, Kniest syndromes, metatrophic dysplasia, vertebra epiphysis metaphyseal dysplasia, nerve Fibromatosis, Legius syndromes, LEOPARD syndromes, Noonan syndromes, hereditary gingival fibromatosis, 1 type nerve are fine Tie up knurl disease, Legius syndromes, Cardiofaciocutaneous syndromes, Costello syndromes, SHOX deficiency diseases, special hair Property of short and small stature, growth hormone deficiency, osteoarthritis, cleidocranial dysostosis, craniosynostosis (such as Muenke synthesis Sign, Crouzon syndromes, Apert syndromes, Jackson-Weiss syndromes, Pfeiffer syndromes or Crouzonodermoskeletal syndromes), refer to/toe type, brachydactylia/toe, camptodactylia/toe, polydactyly, zygodactyly, Dyssegmental depauperations, enchondrosis, fibrous dysplasia, hereditary multiple exostoses, hypophosphate Hypophosphatemic rickets, Jaffe-Lichtenstein syndromes, Marfan syndromes, McCune-Albright syndromes, bone are hard Change disease and osteopoikilosis.

56. the purposes of claim 54, wherein the disease is ophthalmology disease.

57. the purposes of claim 54 or 55, wherein the disease is achondroplasia.

58. apply in Claims 1-4 6 the CNP prodrugs of any one or its pharmaceutically acceptable salt or claim 47 or The method of 48 pharmaceutical composition, wherein methods described include applying CNP prodrugs, its pharmaceutically acceptable salt or drug regimen The step of thing, described apply pass through following progress：Local, enteral or parenteral administration or applications, the side of injection or infusion In method, including intra-articular, periarticular, intracutaneous, subcutaneous, intramuscular, intravenous, bone, in intraperitoneal, intrathecal, intracapsular, socket of the eye, glass Noted in glass body, in tympanum, in bladder, in heart, under transtracheal, subcutaneous, capsule, under arachnoid, in backbone, in intra-ventricle, breastbone Penetrate and be transfused, brain, the direct ventricles of the brain are directly delivered to by the implanted device for allowing for present invention etc. to be transported to brain tissue or brain liquid Interior injection or infusion, brain or brain relevant range are injected or be infused into, be expelled to space under choroid, injected after socket of the eye and eyes drip Note.

59. the CNP prodrugs of any one or its pharmaceutically acceptable salt or claim 47 or 48 in claim 1-46 Pharmaceutical composition, it is used to treat achondroplasia by being subcutaneously injected.